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WO2021033849A1 - Beauveria bassiana culture medium comprising coffee grounds and beauveria bassiana culturing method using same - Google Patents

Beauveria bassiana culture medium comprising coffee grounds and beauveria bassiana culturing method using same Download PDF

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Publication number
WO2021033849A1
WO2021033849A1 PCT/KR2019/017700 KR2019017700W WO2021033849A1 WO 2021033849 A1 WO2021033849 A1 WO 2021033849A1 KR 2019017700 W KR2019017700 W KR 2019017700W WO 2021033849 A1 WO2021033849 A1 WO 2021033849A1
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Prior art keywords
coffee
medium
meal
comparative example
culture
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Ceased
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PCT/KR2019/017700
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French (fr)
Korean (ko)
Inventor
하병덕
김종협
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Microsolution Co Ltd
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Microsolution Co Ltd
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Publication of WO2021033849A1 publication Critical patent/WO2021033849A1/en
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor

Definitions

  • the present invention relates to a Bacillus bacteria culture medium and a method of culturing Bacillus bacteria using the same, and more particularly, to a Bacillus bacteria culture medium including coffee meal and a method of culturing Bacillus bacteria using the same.
  • Bacillus bacteria are parasitic on the bodies of larvae, pupae, and imago of various types of silkworm moths, grasshoppers and cicadas, and white powdery insect hyphae and conidia cover the body of insects.
  • Baekgangjam which is parasitic in silkworm and silkworm pupa, is used as a herbal medicine material for the treatment of stroke and hives. It treats stroke, has excellent effect on ankylosing paralysis and convulsive disease, high blood pressure, and fever. Guanwasa caused by stroke , It has been used for the treatment of children's game, headache, and pulmonary tuberculosis, and has been used for diabetes and acute mastitis.
  • An object of the present invention is to increase production by efficiently culturing Beauveria bassiana , which can be used as an insect control or herbal medicine, and a medium for cultivation of Bacillus bacteria containing coffee meal as a main component that does not require separate grinding or sterilization of the medium material.
  • Another object of the present invention is to provide a method for cultivating Bacillus bacteria capable of remarkably increasing the yield by using a medium for cultivation of Bacillus bacteria containing as a main component coffee meal that does not require pulverization or sterilization treatment.
  • a medium for culturing Baekgang (Beauveria bassiana) containing coffee leaf is provided.
  • the medium based on 100 parts by weight of coffee meal, 0.1 to 0.8 parts by weight of sugar; 0.1 to 0.8 parts by weight of yeast extract powder; And 0.01 to 0.2 parts by weight of a mineral component.
  • the coffee meal may be a coffee meal powder in a pulverized state.
  • the average particle diameter of the coffee foil may be 180 to 220 ⁇ m.
  • the coffee foil may have a specific surface area (BET) of 0.07 to 0.13 m 2 /g.
  • the moisture content of the coffee foil may be adjusted to 30 to 40wt%.
  • the medium containing the coffee leaf based on 100 parts by weight of coffee leaf, 0.1 to 0.8 parts by weight of sugar; 0.1 to 0.8 parts by weight of yeast extract powder; And 0.01 to 0.2 parts by weight of a mineral component.
  • Bacillus bacteria using a medium containing the coffee leaf Beauveria bassiana
  • the cultivation method includes the steps of: (a) extracting the roasted coffee beans with hot water and preparing the remaining coffee meal; (b) Bacillus bacteria (Beauveria ) in the medium containing the coffee leaf bassiana ) the first culture step of inoculating and culturing; (c) preparing a dehumidified medium by dehumidifying the first cultured medium; And (d) a secondary culture step of additionally culturing Bacillus bacteria in the dehumidified medium.
  • the coffee meal of step (a) may be a powder in a finely pulverized state.
  • the primary culture of step (b) may be started in a state in which the water content of the medium is 25 to 35 wt%.
  • the primary culture of step (b) may be performed for 2 to 3 days.
  • step (c) the dehumidification may be performed so that the moisture content of the first cultured medium is 5 to 15 wt%.
  • step (c) the dehumidification may be performed for 0.5 to 3 hours.
  • coffee meal may be additionally added to the medium before or during the second culture.
  • the secondary culture of step (d) may be performed for 2 to 3 days.
  • the medium for cultivation of Bacillus bacteria containing coffee leaf of the present invention is made of coffee leaf as a main component, and coffee leaf is a porous material with very developed micropores, and it has high antibacterial properties due to high polyphenol content, so it is difficult for bacteria to propagate. It is easy to prepare a medium because there is no need to perform a separate pulverization process or sterilize to prevent bacterial growth, and when cultivating white bacterium using such a medium for culturing white bacterium, it is not necessary to use rice bran or almond meal. Compared to the material, it can significantly improve the cultivation efficiency of Bacillus bacteria, enabling mass production, which is advantageous for use as insect control in farmland.
  • Baekgang (Beauveria bassiana ) culture medium of the present invention based on 100 parts by weight of coffee meal, 0.1 to 0.8 parts by weight of saccharides; 0.1 to 0.8 parts by weight of yeast extract powder; And 0.01 to 0.2 parts by weight of a mineral component.
  • the coffee foil is a coffee foil powder in a finely pulverized state.
  • the coffee foil has a specific surface area (BET) of 0.07 to 0.13 m 2 /g, and has a microporous structure, so that a sufficient surface area can be provided even without a separate grinding process.
  • the coffee meal powder may be 180 to 220 ⁇ m, preferably 190 to 210 ⁇ m, and even more preferably 195 to 205 ⁇ m.
  • the moisture content of the coffee foil is preferably adjusted to 30 to 40wt%, more preferably 32 to 38wt%, even more preferably 34 to 36wt%.
  • Baekganggyun (Beauveria) using a medium containing coffee leaf of the present invention bassiana).
  • the roasted coffee beans are extracted with hot water and the remaining coffee leaves are prepared (step a).
  • the coffee leaves may be used at home or at a coffee shop, and may be used regardless of the type of coffee beans.
  • the primary culture is preferably started with a water content of 25 to 35 wt% of the medium, more preferably 27 to 33 wt%, even more preferably 28 to 32 wt%. If it is out of the above range, it is difficult to reach the number of bacteria above a certain level because the initial proliferation power is lowered.
  • the primary culture is preferably performed for 2 to 3 days.
  • the first cultured medium is dehumidified to prepare a dehumidified medium (step c).
  • the dehumidification is preferably performed so that the moisture content of the medium in which the primary culture is completed is 5 to 15 wt%, more preferably 7 to 13 wt%, and more preferably 8 to 12 wt%.
  • the dehumidification is preferably performed for 0.5 to 3 hours, more preferably for 1 to 3 hours, more preferably for 2 to 3 hours.
  • the formation of Bacillus spores may be actively performed, and when the water content of the medium is less than 5wt% or exceeds 15wt%, the formation of Bacillus spores cannot be properly achieved.
  • step d a second culture of Bacillus bacteria is carried out in the dehumidified medium.
  • coffee meal Before or during the secondary culture, coffee meal may be additionally added to the medium, and the coffee meal may be pulverized coffee meal powder similar to the coffee meal used in step (a). ) May be the same as or different.
  • the secondary culture is preferably performed for 2 to 3 days.
  • Example 1 Culture of pulverized coffee meal
  • coffee meal powder having an average particle diameter of about 200 ⁇ m was prepared, and moisture was added to a moisture content of about 35 wt%.
  • coffee meal powder 99, glucose 0.4, yeast extract powder 0.5, minerals (calcium, potassium, MgSO 4 , K 3 PO 4 ) components were mixed in a weight ratio of 99:0.4:0.5:0.1 to prepare a coffee leaf medium .
  • Baekgang (Beauveria bassiana ) KACC 40039 was inoculated with 10% (w/v) in the coffee leaf medium thus prepared and cultured at 25° C. for 2 days. Thereafter, the medium in which Bacillus was cultured was further cultured at 25° C. while dehumidifying so that the moisture content was 10 wt% or less for 2 days.
  • Example 1 Except that the dry coffee meal powder of Example 1 was pulverized and pulverized to an average particle diameter of about 150 ⁇ m, Bacillus bacteria were cultured in the same manner as in Example 1.
  • Bacillus bacteria were cultured in the same manner as in Example 1, except that the coffee meal powder of Example 1 was sterilized with steam for 15 minutes at 120°C and 1.2 atm in an autoclave.
  • Bacillus bacteria were cultured in the same manner as in Example 1, except that the ground coffee meal powder of Example 2 was sterilized as in Example 3 and then used.
  • Example 3 After the coffee meal powder of Example 1 was sterilized as in Example 3, the Bacillus bacteria were cultured in the same manner as in Example 1, except that a material to which a commercially available green tea polyphenol was added as much as 1 wt% of the coffee meal powder was used I did.
  • Bacillus bacteria were cultured in the same manner as in Example 1, except that a material obtained by adding a commercially available green tea polyphenol as much as 1 wt% of the coffee leaf powder to the ground coffee leaf powder of Example 2 was used.
  • Example 7 Grinding + sterilization + cultivation of Bacillus bacteria using polyphenol coffee foil
  • Example 2 In the same manner as in Example 1, except that the ground coffee meal powder of Example 2 was sterilized as in Example 3, and then a material to which a commercially available green tea polyphenol was added as much as 1 wt% of the ground coffee leaf powder was used. Bacillus bacteria were cultured.
  • Bacillus bacteria were cultured in the same manner as in Example 1, except that almond oil was extracted instead of coffee meal powder and a medium was prepared using almond meal powder having an average particle diameter of 250 ⁇ m, which is the remaining residue.
  • Bacillus bacteria were cultured in the same manner as in Example 2, except that the almond meal powder of Comparative Example 1 was pulverized to prepare a medium with a powder having an average particle diameter of 150 ⁇ m.
  • Bacillus bacteria were cultured in the same manner as in Example 3, except that the almond meal powder of Comparative Example 1 was used.
  • Bacillus bacteria were cultured in the same manner as in Example 4, except that the crushed almond meal of Comparative Example 2 was used.
  • Bacillus bacteria were cultured in the same manner as in Example 5, except that the almond meal of Comparative Example 1 was used.
  • Bacillus bacteria were cultured in the same manner as in Example 6, except that the crushed almond meal of Example 2 was used.
  • Bacillus bacteria were cultured in the same manner as in Example 5, except that the crushed almond meal of Comparative Example 2 was used.
  • Bacillus bacteria were cultured in the same manner as in Example 1, except that rice bran powder having an average particle diameter of 200 ⁇ m was used instead of coffee foil.
  • Bae ganglia were cultured in the same manner as in Example 2, except that the rice bran powder of Comparative Example 8 was pulverized to have an average particle diameter of 150 ⁇ m.
  • Comparative Example 8 Bacillus bacteria were cultured in the same manner as in Example 3, except that rice bran powder was used.
  • Bacillus bacteria were cultured in the same manner as in Example 4, except that the ground rice bran of Comparative Example 9 was used.
  • Bacillus bacteria were cultured in the same manner as in Example 5, except that the rice bran powder of Comparative Example 8 was used.
  • Bacillus bacteria were cultured in the same manner as in Example 6, except that the ground rice bran of Comparative Example 9 was used.
  • Bacillus bacteria were cultured in the same manner as in Example 7, except that the ground rice bran of Comparative Example 9 was used.
  • Examples 1 to 7, and Comparative Examples 1 to 14 are summarized in Table 1 below for the culture conditions of Bacillus bacteria.
  • Example 1 Coffee leaf ⁇ ⁇ ⁇ Example 2 ⁇ ⁇ ⁇ Example 3 ⁇ ⁇ ⁇ Example 4 ⁇ ⁇ ⁇ Example 5 ⁇ ⁇ ⁇ Example 6 ⁇ ⁇ ⁇ Example 7 ⁇ ⁇ ⁇ Comparative Example 1 Almond gourd ⁇ ⁇ ⁇ Comparative Example 2 ⁇ ⁇ ⁇ Comparative Example 3 ⁇ ⁇ ⁇ Comparative Example 4 ⁇ ⁇ ⁇ Comparative Example 5 ⁇ ⁇ ⁇ Comparative Example 6 ⁇ ⁇ ⁇ Comparative Example 7 ⁇ ⁇ ⁇ Comparative Example 8 Rice bran ⁇ ⁇ ⁇ Comparative Example 9 ⁇ ⁇ ⁇ Comparative Example 10 ⁇ ⁇ ⁇ Comparative Example 11 ⁇ ⁇ ⁇ Comparative Example 12 ⁇ ⁇ ⁇ Comparative Example 13 ⁇ ⁇ ⁇ Comparative Example 14 ⁇ ⁇ ⁇
  • Example 5 using pulverized coffee foil was 12 times or more compared to Comparative Example 5 using almond meal under the same conditions, and 15 times or more compared to Comparative Example 12 using rice bran It was determined that the strain concentration was high.
  • Examples 5 and 7 using coffee grounds did not show a significant difference in pulverization status and strain concentration. In other words, it was found that when the coffee leaf was used as a medium material, an additional grinding process was not required.
  • Comparative Example 7 using crushed almond meal had a final strain concentration four or more times higher than that of Comparative Example 5 using pulverized almond meal. That is, in the case of using almond meal as a medium material, it was found that the cultivation efficiency of four or more times could be exhibited by performing an additional pulverization process, and it was found that the production of strains could be increased only when the almond meal was pulverized.
  • Comparative Example 12 and Comparative Example 14 using rice bran Comparative Example 14 using pulverized rice bran showed a final strain concentration of 4 times or more compared to Comparative Example 12 using pulverized rice bran. It was found that rice bran can also increase the strain production only by performing an additional grinding process.
  • the medium containing the coffee meal of the present invention not only exhibits significantly higher cultivation efficiency than the case of using a medium containing almond meal or rice bran, which are other waste materials, and the coffee meal has a microporous structure. It can be seen that since the specific surface area is higher than that of gourd or rice bran, there is no need for an additional pulverization process, which can reduce process steps.
  • the concentrations of the strains of the final cultured Bacillus bacteria were compared with those obtained by sterilizing coffee meal, almond meal, or rice bran, and are shown in Table 3. At this time, since the additional pulverization greatly influenced the cultivation efficiency in the almond meal and rice bran, it was considered to be performed uniformly, and the polyphenol was not added.
  • Example 2 using pulverized coffee foil was 17 times or more compared to Comparative Example 2 using almond meal under the same conditions, and 8 times or more compared to Comparative Example 9 using rice bran It was determined that the strain concentration was high.
  • Examples 2 and 4 using coffee foil did not show a significant difference between whether or not sterilized and the strain concentration. In other words, it was found that the sterilization treatment process was not required when coffee leaf was used as a medium material.
  • Comparative Example 2 using sterilized almond meal had twice the final strain concentration as compared to Comparative Example 4 using almond meal that was not sterilized. Appeared to be high. That is, in the case of using almond meal as a medium material, it was found that the cultivation efficiency of more than two times could be exhibited by performing sterilization treatment, and it was found that the production of strains could be increased only when almond meal was sterilized. In addition, when comparing Comparative Example 9 and Comparative Example 11 using rice bran, there was no significant difference in the final strain concentration. Rice bran, unlike almond meal, did not require sterilization.
  • the medium containing the coffee meal of the present invention not only exhibits significantly higher cultivation efficiency than the case of using a medium containing almond meal or rice bran, which is a waste material, but also sterilizes because a large amount of polyphenol is contained in the coffee leaf. It can be seen that there is no need for treatment, which has the effect of reducing process steps.
  • Example 4 Coffee leaf ⁇ ⁇ ⁇ 1.03 ⁇ 10 9
  • Example 4 ⁇ ⁇ ⁇ 1.09 ⁇ 10 9
  • Example 6 ⁇ ⁇ ⁇ 1.12 ⁇ 10 9
  • Example 7 ⁇ ⁇ ⁇ 1.07 ⁇ 10 9
  • Comparative Example 2 Almond gourd ⁇ ⁇ ⁇ 3.80 ⁇ 10 7
  • Comparative Example 4 ⁇ ⁇ ⁇ 1.15 ⁇ 10 8
  • Comparative Example 6 ⁇ ⁇ ⁇ 8.22 ⁇ 10 7
  • Comparative Example 9 Rice bran ⁇ ⁇ ⁇ 1.26 ⁇ 10 8
  • Comparative Example 11 ⁇ ⁇ ⁇ 1.13 ⁇ 10 8
  • Comparative Example 13 ⁇ ⁇ ⁇ 1.03 ⁇ 10 8 Comparative Example 14 ⁇ ⁇ ⁇ 1.26 ⁇ 10 8
  • Examples 2, 4, 6 and 7 of the present invention when comparing the cultivation efficiency according to the medium material, are 10 times to 10 times compared to Comparative Examples using almond meal or rice bran under the same conditions. The culture efficiency of the medium material was remarkably high by 30 times.
  • the strain concentration in Examples 2, 4, 6, and 7 using coffee ground did not show a significant difference. That is, it can be seen that in the case of using coffee leaf as a medium material, addition of polyphenol does not have a special effect on the strain concentration.
  • the medium containing the coffee meal of the present invention not only exhibits significantly higher cultivation efficiency than the case of using a medium containing almond meal or rice bran, which is a waste material, but also sterilizes because a large amount of polyphenol is contained in the coffee leaf. It can be seen that there is no need for treatment, which has the effect of reducing process steps.
  • Example 1 Coffee leaf ⁇ ⁇ ⁇ 1.13 ⁇ 10 9 Comparative Example 1 Almond gourd ⁇ ⁇ ⁇ 2.22 ⁇ 10 7 Comparative Example 8 Rice bran ⁇ ⁇ ⁇ 3.45 ⁇ 10 7
  • the final strain concentration of Example 1 using coffee meal was about 30 times as compared to Comparative Example 1 using almond meal, and about 100 times as compared to Comparative Example 8 using rice bran.
  • the medium for cultivation of Bacillus bacteria containing coffee foil of the present invention can exhibit remarkably high cultivation efficiency of Bacillus bacteria without additional processes such as pulverization or sterilization, or addition of ingredients for sterilization by using coffee leaf as a main material.
  • the medium for cultivation of Bacillus bacteria containing coffee leaf of the present invention is made of coffee leaf as a main component, and coffee leaf is a porous material with very developed micropores, and it has high antibacterial properties due to high polyphenol content, so it is difficult for bacteria to propagate. It is easy to prepare a medium because there is no need to perform a separate pulverization process or sterilize to prevent bacterial growth, and when cultivating white bacterium using such a medium for culturing white bacterium, it is not necessary to use rice bran or almond meal. Compared to the material, it can significantly improve the cultivation efficiency of Bacillus bacteria, enabling mass production, which is advantageous for use as insect control in farmland.

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Abstract

The present invention relates to a Beauveria bassiana culture medium comprising coffee grounds and a Beauveria bassiana culturing method using same. Since coffee grounds are used for the main material, the present invention does not require separate grinding or sterilization of a medium material, and thus the medium can be prepared easily and the culture yield of the Beauveria bassiana can be notably increased.

Description

커피박을 포함하는 백강균 배양용 배지 및 이를 이용한 백강균의 배양방법Bacillus cultivation medium containing coffee leaf and method for cultivating white bacterium using the same

본 발명은 백강균 배양 배지 및 이를 이용한 백강균의 배양방법에 관한 것으로, 더욱 상세하게는 커피박을 포함하는 백강균 배양 배지 및 이를 이용한 백강균의 배양방법에 관한 것이다.The present invention relates to a Bacillus bacteria culture medium and a method of culturing Bacillus bacteria using the same, and more particularly, to a Bacillus bacteria culture medium including coffee meal and a method of culturing Bacillus bacteria using the same.

백강균(Beauveria bassiana)을 대상으로한 본격적인 연구는 1956년 적합한 배양법을 제시한 Muller-Kogler에 의해서 시작되었다. 동충하초(Cordyceps)로 알려진 곤충 병원성 균주(Entonopathogenic fungi) 중 불완전균류인 백강균(Beauveria bassiana)은 곤충에 굳음병을 일으키는 균으로 메뚜기목, 나비목, 딱정벌레목 등 넓은 기주 범위를 가지고 환경 변화에 강한 종으로 알려져 있다. 주로 곤충 표피를 통하여 내부로 침입하여 곤충의 면역작용을 차단하거나 독성물질을 분비하여 기주를 치사시킨다. 백강균은 여러 종류의 누에나방, 메뚜기과, 매미과 등의 유충, 번데기, 성충의 몸에 기생하여 백색 분말상의 곤충 균사와 분생 포자가 곤충의 몸체를 뒤덮는데 생육시에는 백색이나 건조하면 회황색이 된다. 누에와 누에 번데기에 기생한 백강균을 백강잠(Bombyx mori)이라고 하는데 중풍, 두드러기 등의 치료에 한약 재료로 이용되며 뇌졸중을 치료하며 강직성 마비 및 경련 질환, 고혈압, 해열에 효력이 탁월하며 중풍으로 인한 구안와사, 어린아이의 경기치료, 두통의 치료, 폐결핵의 치료 등에 이용되어 왔으며 당뇨병 및 급선 유선염 등에 이용된 예가 있다. 식물 병원성 세균 중 뽕나무의 줄기썩음병을 발생시키는 에르위니아 라폰티시(Erwinia rhapontici), 세균성 오갈병의 슈도모나스 시린개(Pseudomonas syringae), 가지 썩음병의 원인균인 잔토모나스 캄페스트리스(Xantomonas campestris)를 백강균 J33, J34, J54, J57, J157, J200 총 6 균주 배양액으로 항균활성을 측정한 결과, 백강균 J200에서만 뽕나무병원성세균에 대한 가장 우수한 항균활성을 보였으며 같은 종이라도 균주에 따라 항균활성의 차이를 나타냈다고 보고하였다. 다양한 종류의 해충을 치사시키는 특성을 가지고 있어 백강균을 포함한 곤충 병원성 균주의 식중독균에 대한 항균활성을 보고하는 등 천연물질인 백강균에 관한 관심이 증가하고 있는 실정이다. 또한, 백강균(KACC 40024, KACC 40039, KACC 40377, HYI) 균주의 거세미나방(Agrotis segetum)에 대한 살충력을 검정한 결과 KACC 40039, 백강균 HYI이 높은 살충력을 나타내어 거세미나방 방제용 미생물 살충제로의 가능성이 나타났다고 보고하였다. 이와 같이 한약재료 또는 곤충 방제용으로 사용되는 백강균을 간편한 생산공정으로 효율적으로 생산할 수 있는 배양방법이 필요한 실정이다. 한편, 커피 원두에서 커피액을 추출하고 남은 잔사인 커피박은 세계적으로 연간 6백만 톤 이상 폐기물로 다량 발생하고 있으나 이에 대한 활용 및 처리방안이 부족한 상황이다. 따라서 커피박의 재활용에 대한 연구가 활발히 이루어지고 있으며, 커피박을 연료, 사료 또는 비료 첨가물로 사용하거나, 버섯 생산을 위한 기질 및 바이오디젤 원료로 활용하는 것에 대한 연구가 이루어지고 있다. A full-fledged study on Beauveria bassiana began in 1956 by Muller-Kogler, who suggested an appropriate culture method. Among the entonopathogenic fungi known as Cordyceps, Beauveria bassiana, an incomplete fungus, is a fungus that causes stiffness in insects.It has a wide host range, such as the Locust Order, Lepidoptera, and Beetle Order, and is a species resistant to environmental changes. Is known. Mainly through the insect epidermis, it invades the inside to block the insect's immune function or secrete toxic substances to kill the host. Bacillus bacteria are parasitic on the bodies of larvae, pupae, and imago of various types of silkworm moths, grasshoppers and cicadas, and white powdery insect hyphae and conidia cover the body of insects. Baekgangjam (Bombyx mori), which is parasitic in silkworm and silkworm pupa, is used as a herbal medicine material for the treatment of stroke and hives. It treats stroke, has excellent effect on ankylosing paralysis and convulsive disease, high blood pressure, and fever. Guanwasa caused by stroke , It has been used for the treatment of children's game, headache, and pulmonary tuberculosis, and has been used for diabetes and acute mastitis. Among plant pathogenic bacteria, Erwinia rhapontici, which causes stem rot of mulberry trees, Pseudomonas syringae, and Xantomonas campestris, which is the causative agent of eggplant rot, were treated with Bacillus J33, As a result of measuring antimicrobial activity with a total of 6 strains of J34, J54, J57, J157, J200, Baekgang bacteria J200 showed the best antibacterial activity against Morus pathogenic bacteria, and even the same species showed differences in antibacterial activity according to strains. I did. Since it has the property of killing various types of pests, interest in the natural substance Baekgang bacillus is increasing, such as reporting the antibacterial activity against food poisoning bacteria of insect pathogenic strains including Bacillus bacillus. In addition, as a result of testing the insecticidal power of Agrotis segetum strains of Bacillus (KACC 40024, KACC 40039, KACC 40377, HYI) strains, KACC 40039 and Bacillus HYI showed high insecticidal power, and thus became a microbial insecticide for controlling castor moths. It reported that a possibility appeared. As such, there is a need for a cultivation method that can efficiently produce Bacillus bacillus, which is used for herbal medicine or insect control, through a simple production process. On the other hand, coffee meal, the residue left after extracting coffee liquid from coffee beans, is generated in large quantities as wastes of more than 6 million tons per year in the world, but there is a lack of utilization and disposal plans for this. Therefore, research on the recycling of coffee meal is being actively conducted, and studies on using coffee meal as a fuel, feed or fertilizer additive, or as a substrate and biodiesel raw material for mushroom production are being conducted.

본 발명의 목적은 곤충방제나 한약재 등으로 사용될 수 있는 백강균(Beauveria bassiana)을 효율적으로 배양하여 생산량을 높이고, 배지 재료를 별도로 분쇄하거나 살균 처리가 필요없는 커피박을 주성분으로 포함하는 백강균 배양용 배지를 제공하는 데 있다. 본 발명의 다른 목적은 배지 재료를 별도로 분쇄하거나 살균 처리가 필요없는 커피박을 주성분으로 포함하는 백강균 배양용 배지를 이용하여 생산량을 현저히 높일 수 있는 백강균의 배양방법을 제공하는 데 있다. An object of the present invention is to increase production by efficiently culturing Beauveria bassiana , which can be used as an insect control or herbal medicine, and a medium for cultivation of Bacillus bacteria containing coffee meal as a main component that does not require separate grinding or sterilization of the medium material. To provide. Another object of the present invention is to provide a method for cultivating Bacillus bacteria capable of remarkably increasing the yield by using a medium for cultivation of Bacillus bacteria containing as a main component coffee meal that does not require pulverization or sterilization treatment.

본 발명의 일 측면에 따르면, 커피박을 포함하는 백강균(Beauveria bassiana) 배양용 배지가 제공된다.According to one aspect of the present invention, a medium for culturing Baekgang (Beauveria bassiana) containing coffee leaf is provided.

상기 배지는, 커피박 100중량부를 기준으로, 당류 0.1 내지 0.8중량부; 효모추출분말 0.1 내지 0.8중량부; 및 미네랄 성분 0.01 내지 0.2중량부;를 포함할 수 있다.The medium, based on 100 parts by weight of coffee meal, 0.1 to 0.8 parts by weight of sugar; 0.1 to 0.8 parts by weight of yeast extract powder; And 0.01 to 0.2 parts by weight of a mineral component.

상기 커피박은 미분쇄 상태의 커피박 분말일 수 있다.The coffee meal may be a coffee meal powder in a pulverized state.

상기 커피박의 평균입경은 180 내지 220㎛ 일 수 있다.The average particle diameter of the coffee foil may be 180 to 220㎛.

상기 커피박은 비표면적(BET)이 0.07 내지 0.13 m2 /g 일 수 있다.The coffee foil may have a specific surface area (BET) of 0.07 to 0.13 m 2 /g.

상기 커피박의 수분함량은 30 내지 40wt% 로 조절된 것일 수 있다.The moisture content of the coffee foil may be adjusted to 30 to 40wt%.

본 발명의 다른 하나의 측면에 따르면, 커피박을 포함하는 배지를 이용한 백강균(Beauveria bassiana)의 배양방법이 제공된다.According to another aspect of the present invention, there is provided a method of culturing Baekgang (Beauveria bassiana) using a medium containing coffee meal.

상기 커피박을 포함하는 배지는, 커피박 100중량부를 기준으로, 당류 0.1 내지 0.8중량부; 효모추출분말 0.1 내지 0.8중량부; 및 미네랄 성분 0.01 내지 0.2중량부;를 포함할 수 있다.The medium containing the coffee leaf, based on 100 parts by weight of coffee leaf, 0.1 to 0.8 parts by weight of sugar; 0.1 to 0.8 parts by weight of yeast extract powder; And 0.01 to 0.2 parts by weight of a mineral component.

상기 커피박을 포함하는 배지를 이용한 백강균(Beauveria bassiana)의 배양방법은, (a) 로스팅된 커피 원두를 열수 추출한 후 남은 커피박을 준비하는 단계; (b) 상기 커피박을 포함하는 배지에 백강균(Beauveria bassiana)을 접종하여 배양하는 1차 배양 단계; (c) 상기 1차 배양된 배지를 제습하여 제습된 배지를 준비하는 단계; 및 (d) 상기 제습된 배지에 백강균을 추가로 배양하는 2차 배양 단계;를 포함할 수 있다.Bacillus bacteria using a medium containing the coffee leaf ( Beauveria bassiana ), the cultivation method includes the steps of: (a) extracting the roasted coffee beans with hot water and preparing the remaining coffee meal; (b) Bacillus bacteria (Beauveria ) in the medium containing the coffee leaf bassiana ) the first culture step of inoculating and culturing; (c) preparing a dehumidified medium by dehumidifying the first cultured medium; And (d) a secondary culture step of additionally culturing Bacillus bacteria in the dehumidified medium.

단계 (a)의 상기 커피박은 미분쇄 상태의 분말일 수 있다.The coffee meal of step (a) may be a powder in a finely pulverized state.

단계 (b)의 1차 배양은 상기 배지의 수분함량이 25 내지 35wt%인 상태에서 시작할 수 있다.The primary culture of step (b) may be started in a state in which the water content of the medium is 25 to 35 wt%.

단계 (b)의 1차 배양은 2 내지 3일 동안 수행될 수 있다.The primary culture of step (b) may be performed for 2 to 3 days.

단계 (c)에서 상기 제습은 1차 배양된 상기 배지의 수분함량이 5 내지 15wt%가 되도록 수행할 수 있다.In step (c), the dehumidification may be performed so that the moisture content of the first cultured medium is 5 to 15 wt%.

단계 (c)에서 상기 제습은 0.5 내지 3시간 동안 수행할 수 있다.In step (c), the dehumidification may be performed for 0.5 to 3 hours.

단계 (d)에서 상기 2차 배양 전 또는 배양 중 배지에 커피박을 추가 투입할 수 있다.In step (d), coffee meal may be additionally added to the medium before or during the second culture.

단계 (d)의 2차 배양은 2 내지 3일 동안 수행될 수 있다.The secondary culture of step (d) may be performed for 2 to 3 days.

본 발명의 커피박을 포함하는 백강균 배양용 배지는 커피박을 주성분으로 하고, 커피박은 미세 기공이 매우 발달한 다공성 재료이며, 폴리페놀 함량이 높아 항균성이 있어 세균이 번식하기 어려우므로 표면적으로 넓혀주기 위해 별도로 분쇄공정을 수행하거나, 세균 번식을 방지하기 위하여 살균처리를 할 필요가 없어 배지 제조가 용이하고, 이와 같은 백강균 배양용 배지를 이용하여 백강균을 배양할 경우 종래 사용되는 미강이나 아몬드박과 같은 재료에 비해 백강균 배양 효율을 현저히 향상시킬 수 있어 대량생산을 가능케 하여 농지에서의 곤충방제로 활용하기에 유리하다.The medium for cultivation of Bacillus bacteria containing coffee leaf of the present invention is made of coffee leaf as a main component, and coffee leaf is a porous material with very developed micropores, and it has high antibacterial properties due to high polyphenol content, so it is difficult for bacteria to propagate. It is easy to prepare a medium because there is no need to perform a separate pulverization process or sterilize to prevent bacterial growth, and when cultivating white bacterium using such a medium for culturing white bacterium, it is not necessary to use rice bran or almond meal. Compared to the material, it can significantly improve the cultivation efficiency of Bacillus bacteria, enabling mass production, which is advantageous for use as insect control in farmland.

본 발명은 다양한 변환을 가할 수 있고 여러 가지 실시예를 가질 수 있는 바, 특정 실시예들을 도면에 예시하고 상세한 설명에 상세하게 설명하고자 한다. 그러나, 이는 본 발명을 특정한 실시 형태에 대해 한정하려는 것이 아니며, 본 발명의 사상 및 기술 범위에 포함되는 모든 변환, 균등물 내지 대체물을 포함하는 것으로 이해되어야 한다. 본 발명을 설명함에 있어서 관련된 공지 기술에 대한 구체적인 설명이 본 발명의 요지를 흐릴 수 있다고 판단되는 경우 그 상세한 설명을 생략한다. 본 출원에서 사용한 용어는 단지 특정한 실시예를 설명하기 위해 사용된 것으로, 본 발명을 한정하려는 의도가 아니다. 단수의 표현은 문맥상 명백하게 다르게 뜻하지 않는 한, 복수의 표현을 포함한다. 본 출원에서, "포함하다" 또는 "가지다" 등의 용어는 명세서상에 기재된 특징, 숫자, 단계, 동작, 구성요소, 부품 또는 이들을 조합한 것이 존재함을 지정하려는 것이지, 하나 또는 그 이상의 다른 특징들이나 숫자, 단계, 동작, 구성요소, 부품 또는 이들을 조합한 것들의 존재 또는 부가 가능성을 미리 배제하지 않는 것으로 이해되어야 한다.Since the present invention can apply various transformations and have various embodiments, specific embodiments are illustrated in the drawings and will be described in detail in the detailed description. However, this is not intended to limit the present invention to a specific embodiment, it is to be understood to include all conversions, equivalents, and substitutes included in the spirit and scope of the present invention. In describing the present invention, when it is determined that a detailed description of a related known technology may obscure the subject matter of the present invention, a detailed description thereof will be omitted. The terms used in the present application are only used to describe specific embodiments, and are not intended to limit the present invention. Singular expressions include plural expressions unless the context clearly indicates otherwise. In the present application, terms such as "comprise" or "have" are intended to designate the presence of features, numbers, steps, actions, components, parts, or combinations thereof described in the specification, but one or more other features. It is to be understood that the presence or addition of elements or numbers, steps, actions, components, parts, or combinations thereof, does not preclude in advance.

이하, 본 발명의 백강균(Beauveria bassiana) 배양용 배지에 대해 설명하도록 한다.Hereinafter, a description will be given of the culture medium for Baekgang (Beauveria bassiana ) culture of the present invention.

본 발명의 백강균(Beauveria bassiana) 배양용 배지는 커피박 100중량부를 기준으로, 당류 0.1 내지 0.8중량부; 효모추출분말 0.1 내지 0.8중량부; 및 미네랄 성분 0.01 내지 0.2중량부;를 포함할 수 있다. Baekgang (Beauveria bassiana ) culture medium of the present invention, based on 100 parts by weight of coffee meal, 0.1 to 0.8 parts by weight of saccharides; 0.1 to 0.8 parts by weight of yeast extract powder; And 0.01 to 0.2 parts by weight of a mineral component.

상기 커피박은 미분쇄 상태의 커피박 분말상기 커피박은 비표면적(BET)이 0.07 내지 0.13 m2 /g 이며, 미세 다공성 구조이기 때문에 별도로 분쇄 과정을 거치지 않더라도 충분한 표면적을 제공할 수 있다. The coffee foil is a coffee foil powder in a finely pulverized state. The coffee foil has a specific surface area (BET) of 0.07 to 0.13 m 2 /g, and has a microporous structure, so that a sufficient surface area can be provided even without a separate grinding process.

상기 커피박 분말은 180 내지 220㎛ 일 수 있고, 바람직하게는 190 내지 210㎛, 더욱 더 바람직하게는 195 내지 205㎛ 일 수 있다.The coffee meal powder may be 180 to 220 μm, preferably 190 to 210 μm, and even more preferably 195 to 205 μm.

상기 커피박의 수분함량은 30 내지 40wt%로 조절된 것이 바람직하고, 더욱 바람직하게는 32 내지 38wt%, 더욱 더 바람직하게는 34 내지 36wt% 일 수 있다.The moisture content of the coffee foil is preferably adjusted to 30 to 40wt%, more preferably 32 to 38wt%, even more preferably 34 to 36wt%.

이하, 본 발명의 커피박을 포함하는 배지를 이용한 백강균(Beauveria bassiana)의 배양방법에 대해 설명하도록 한다. Hereinafter, Baekganggyun (Beauveria) using a medium containing coffee leaf of the present invention bassiana).

상기 커피박을 포함하는 배지는 앞서 설명한 바와 동일하므로 구체적인 내용은 그 부분을 참조하기로 한다.Since the medium containing the coffee leaf is the same as described above, the detailed information will be referred to that part.

먼저, 로스팅된 커피 원두를 열수 추출한 후 남은 커피박을 준비한다(단계 a). First, the roasted coffee beans are extracted with hot water and the remaining coffee leaves are prepared (step a).

로스팅된 커피 원두를 열수 추출한 후 남은 커피박을 준비한다.After the roasted coffee beans are extracted with hot water, the remaining coffee leaves are prepared.

상기 커피박은 가정이나 커피점에서 배출된 것은 모두 사용할 수 있으며, 원두의 종류에 관계없이 모두 사용할 수 있다.The coffee leaves may be used at home or at a coffee shop, and may be used regardless of the type of coffee beans.

다음으로, 상기 Next, above 커피박을Coffee leaf 포함하는 배지에 백강균( Bacillus bacteria ( BeauveriaBeauveria bassianabassiana )을 접종하여 1차 배양한다(단계 b).) To inoculate the primary culture (step b).

1차 배양은 상기 배지의 수분함량이 25 내지 35wt%인 상태에서 시작하는 것이 바람직하고, 더욱 바람직하게는 27 내지 33wt%, 더욱 더 바람직하게는 28 내지 32wt%의 함량으로 시작하는 것이 바람직하다. 상기 범위를 벗어나는 경우에는 초기증식력이 저하되어 일정 수준 이상의 균수에 도달하기 어렵다.The primary culture is preferably started with a water content of 25 to 35 wt% of the medium, more preferably 27 to 33 wt%, even more preferably 28 to 32 wt%. If it is out of the above range, it is difficult to reach the number of bacteria above a certain level because the initial proliferation power is lowered.

상기 1차 배양은 2 내지 3일 동안 수행하는 것이 바람직하다.The primary culture is preferably performed for 2 to 3 days.

이후, 상기 1차 배양된 배지를 제습하여 제습된 배지를 준비한다(단계 c).Thereafter, the first cultured medium is dehumidified to prepare a dehumidified medium (step c).

상기 제습은 1차 배양이 완료된 배지의 수분함량이 5 내지 15wt%가 되도록 수행하는 것이 바람직하고, 더욱 바람직하게는 7 내지 13wt%, 더욱 바람직하게는 8 내지 12wt% 가 되도록 제습할 수 있다.The dehumidification is preferably performed so that the moisture content of the medium in which the primary culture is completed is 5 to 15 wt%, more preferably 7 to 13 wt%, and more preferably 8 to 12 wt%.

상기 제습은 0.5 내지 3시간 동안 수행하는 것이 바람직하고, 더욱 바람직하게는 1 내지 3시간, 더욱 바람직하게는 2 내지 3시간 동안 수행할 수 있다.The dehumidification is preferably performed for 0.5 to 3 hours, more preferably for 1 to 3 hours, more preferably for 2 to 3 hours.

이와 같은 조건으로 제습된 배지에서 백강균 포자의 형성이 활발히 이루어질 수 있으며, 배지의 수분함량이 5wt% 미만이거나, 15wt%를 초과하는 경우 백강균 포자의 형성이 제대로 이루어질 수 없다.In the dehumidified medium under such conditions, the formation of Bacillus spores may be actively performed, and when the water content of the medium is less than 5wt% or exceeds 15wt%, the formation of Bacillus spores cannot be properly achieved.

다음으로, 상기 제습된 배지에 백강균을 2차 배양한다(단계 d).Next, a second culture of Bacillus bacteria is carried out in the dehumidified medium (step d).

상기 2차 배양 전 또는 배양 중 배지에 커피박을 추가 투입할 수 있으며, 상기 커피박은 단계 (a)에서 사용한 커피박과 마찬가지로 미분쇄 커피박 분말일 수 있으며, 커피박의 원두 종류는 단계 (a)와 동일하거나 상이할 수 있다.Before or during the secondary culture, coffee meal may be additionally added to the medium, and the coffee meal may be pulverized coffee meal powder similar to the coffee meal used in step (a). ) May be the same as or different.

상기 2차 배양은 2 내지 3일간 수행되는 것이 바람직하다.The secondary culture is preferably performed for 2 to 3 days.

실시예 1: 미분쇄 커피박 배양Example 1: Culture of pulverized coffee meal

아라비카종과 로브스타종이 7:3 비율로 혼합된 원두를 고온 및 고압에서 열수추출한 후 남은 평균입경 약 200㎛인 건조 커피박 분말을 준비하여 수분함량이 약 35wt%가 되도록 수분을 가하였다. 이에 따라 얻은 커피박 분말 99, 포도당 0.4, 효모추출분말 0.5, 미네랄(칼슘, 칼륨, MgSO4, K3PO4) 성분이 99:0.4:0.5:0.1의 중량비로 혼합된 커피박 배지를 제조하였다. 이와 같이 제조된 커피박 배지에 백강균(Beauveria bassiana) KACC 40039를 10%(w/v)로 접종하여 25℃에서 2일 동안 배양하였다. 이후, 백강균이 배양된 배지를 2일 동안 수분함량 10wt% 이하가 되도록 제습하면서 25℃에서 추가 배양하였다.After hot water extraction of coffee beans mixed with Arabica species and Lobster species in a ratio of 7:3 at high temperature and high pressure, dry coffee meal powder having an average particle diameter of about 200 μm was prepared, and moisture was added to a moisture content of about 35 wt%. Thus obtained coffee meal powder 99, glucose 0.4, yeast extract powder 0.5, minerals (calcium, potassium, MgSO 4 , K 3 PO 4 ) components were mixed in a weight ratio of 99:0.4:0.5:0.1 to prepare a coffee leaf medium . Baekgang (Beauveria bassiana ) KACC 40039 was inoculated with 10% (w/v) in the coffee leaf medium thus prepared and cultured at 25° C. for 2 days. Thereafter, the medium in which Bacillus was cultured was further cultured at 25° C. while dehumidifying so that the moisture content was 10 wt% or less for 2 days.

실시예 2: 분쇄 커피박 배양Example 2: Culture of ground coffee leaf

실시예 1의 건조 커피박 분말을 분쇄하여 평균입경 약 150㎛로 분말화하여 사용한 것을 제외하고는 실시예 1과 동일한 방법으로 백강균을 배양하였다.Except that the dry coffee meal powder of Example 1 was pulverized and pulverized to an average particle diameter of about 150 µm, Bacillus bacteria were cultured in the same manner as in Example 1.

실시예Example 3: 멸균 3: sterilization 커피박Coffee leaf 배양 culture

실시예 1의 커피박 분말을 오토클레이브에서 120℃, 1.2기압에서 15분 동안 증기로 멸균 처리하여 사용한 것을 제외하고는 실시예 1과 동일한 방법으로 백강균을 배양하였다.Bacillus bacteria were cultured in the same manner as in Example 1, except that the coffee meal powder of Example 1 was sterilized with steam for 15 minutes at 120°C and 1.2 atm in an autoclave.

실시예Example 4: 분쇄 + 멸균 4: grinding + sterilization 커피박Coffee leaf 배양 culture

실시예 2의 분쇄 커피박 분말을 실시예 3과 같이 멸균 처리한 후 사용한 것을 제외하고는 실시예 1과 동일한 방법으로 백강균을 배양하였다.Bacillus bacteria were cultured in the same manner as in Example 1, except that the ground coffee meal powder of Example 2 was sterilized as in Example 3 and then used.

실시예 5: 멸균 + 폴리페놀 추가 커피박 배양Example 5: Sterilization + Polyphenol Addition Coffee Leaf Culture

실시예 1의 커피박 분말을 실시예 3과 같이 멸균 처리한 후, 커피박 분말의 1wt% 만큼 시판하는 녹차 폴리페놀을 추가한 재료를 사용한 것을 제외하고는 실시예 1과 동일한 방법으로 백강균을 배양하였다.After the coffee meal powder of Example 1 was sterilized as in Example 3, the Bacillus bacteria were cultured in the same manner as in Example 1, except that a material to which a commercially available green tea polyphenol was added as much as 1 wt% of the coffee meal powder was used I did.

실시예 6: 분쇄 + 폴리페놀 추가 커피박 배양Example 6: Grinding + Polyphenol Addition Coffee Gourd Culture

실시예 2의 분쇄 커피박 분말에 커피박 분말의 1wt% 만큼 시판하는 녹차 폴리페놀을 추가한 재료를 사용한 것을 제외하고는 실시예 1과 동일한 방법으로 백강균을 배양하였다.Bacillus bacteria were cultured in the same manner as in Example 1, except that a material obtained by adding a commercially available green tea polyphenol as much as 1 wt% of the coffee leaf powder to the ground coffee leaf powder of Example 2 was used.

실시예 7: 분쇄 + 멸균 + 폴리페놀 커피박 이용 백강균 배양Example 7: Grinding + sterilization + cultivation of Bacillus bacteria using polyphenol coffee foil

실시예 2의 분쇄 커피박 분말을 실시예 3과 같이 멸균 처리한 후, 분쇄 커피박 분말의 1wt% 함량만큼 시판하는 녹차 폴리페놀을 추가한 재료를 사용한 것을 제외하고는 실시예 1과 동일한 방법으로 백강균을 배양하였다.In the same manner as in Example 1, except that the ground coffee meal powder of Example 2 was sterilized as in Example 3, and then a material to which a commercially available green tea polyphenol was added as much as 1 wt% of the ground coffee leaf powder was used. Bacillus bacteria were cultured.

비교예 1: 미분쇄 아몬드박 배양Comparative Example 1: Culture of finely ground almond meal

커피박 분말 대신에 아몬드유를 추출하고 남은 찌꺼기인 평균입경 250㎛인 아몬드박 분말을 사용하여 배지를 제조한 것을 제외하고는 실시예 1과 동일한 방법으로 백강균을 배양하였다.Bacillus bacteria were cultured in the same manner as in Example 1, except that almond oil was extracted instead of coffee meal powder and a medium was prepared using almond meal powder having an average particle diameter of 250 μm, which is the remaining residue.

비교예 2: 분쇄 아몬드박 배양Comparative Example 2: Cultivation of ground almond meal

비교예 1의 아몬드박 분말을 분쇄하여 평균입경 150㎛인 분말로 배지를 제조한 것을 제외하고는 실시예 2와 동일한 방법으로 백강균을 배양하였다.Bacillus bacteria were cultured in the same manner as in Example 2, except that the almond meal powder of Comparative Example 1 was pulverized to prepare a medium with a powder having an average particle diameter of 150 μm.

비교예 3: 멸균 아몬드박 배양Comparative Example 3: Sterile almond meal culture

비교예 1의 아몬드박 분말을 사용한 것을 제외하고는 실시예 3과 동일한 방법으로 백강균을 배양하였다.Bacillus bacteria were cultured in the same manner as in Example 3, except that the almond meal powder of Comparative Example 1 was used.

비교예 4: 분쇄 + 멸균 아몬드박 배양Comparative Example 4: Crushed + sterilized almond meal culture

비교예 2의 분쇄 아몬드박을 사용한 것을 제외하고는 실시예 4와 동일한 방법으로 백강균을 배양하였다.Bacillus bacteria were cultured in the same manner as in Example 4, except that the crushed almond meal of Comparative Example 2 was used.

비교예 5: 멸균 + 폴리페놀 추가 아몬드박 배양Comparative Example 5: Sterilization + polyphenol addition almond meal culture

비교예 1의 아몬드박을 사용한 것을 제외하고는 실시예 5와 동일한 방법으로 백강균을 배양하였다. Bacillus bacteria were cultured in the same manner as in Example 5, except that the almond meal of Comparative Example 1 was used.

실시예 6: 분쇄 + 폴리페놀 추가 아몬드박 배양Example 6: Crushing + polyphenol addition almond meal culture

실시예 2의 분쇄 아몬드박을 사용한 것을 제외하고는 실시예 6과 동일한 방법으로 백강균을 배양하였다.Bacillus bacteria were cultured in the same manner as in Example 6, except that the crushed almond meal of Example 2 was used.

비교예 7: 분쇄 + 멸균 + 폴리페놀 추가 아몬드박 배양Comparative Example 7: Crushing + sterilization + polyphenol addition almond meal culture

비교예 2의 분쇄 아몬드박을 사용한 것을 제외하고는 실시예 5와 동일한 방법으로 백강균을 배양하였다.Bacillus bacteria were cultured in the same manner as in Example 5, except that the crushed almond meal of Comparative Example 2 was used.

비교예 8: 미분쇄 미강 배양Comparative Example 8: Culture of pulverized rice bran

커피박 대신에 평균입경 200㎛인 미강 분말을 사용한 것을 제외하고는 실시예 1과 동일한 방법으로 백강균을 배양하였다.Bacillus bacteria were cultured in the same manner as in Example 1, except that rice bran powder having an average particle diameter of 200 μm was used instead of coffee foil.

비교예 9: 분쇄 미강 배양Comparative Example 9: Crushed rice bran culture

비교예 8의 미강 분말을 분쇄하여 평균입경 150㎛로 분말화하여 사용한 것을 제외하고는 실시예 2와 동일한 방법으로 백강균을 배양하였다.Bae ganglia were cultured in the same manner as in Example 2, except that the rice bran powder of Comparative Example 8 was pulverized to have an average particle diameter of 150 μm.

비교예 10: 멸균 미강 배양Comparative Example 10: Sterile rice bran culture

비교예 8 미강 분말을 사용한 것을 제외하고는 실시예 3과 동일한 방법으로 백강균을 배양하였다.Comparative Example 8 Bacillus bacteria were cultured in the same manner as in Example 3, except that rice bran powder was used.

비교예 11: 분쇄 + 멸균 미강 배양Comparative Example 11: Crushing + sterile rice bran culture

비교예 9의 분쇄 미강을 사용한 것을 제외하고는 실시예 4와 동일한 방법으로 백강균을 배양하였다.Bacillus bacteria were cultured in the same manner as in Example 4, except that the ground rice bran of Comparative Example 9 was used.

비교예 12: 멸균 + 폴리페놀 추가 미강 배양Comparative Example 12: Sterilization + polyphenol addition rice cultivation

비교예 8의 미강 분말을 사용한 것을 제외하고는 실시예 5와 동일한 방법으로 백강균을 배양하였다.Bacillus bacteria were cultured in the same manner as in Example 5, except that the rice bran powder of Comparative Example 8 was used.

비교예 13: 분쇄Comparative Example 13: Crushing + 폴리페놀 추가 미강 배양+ Add polyphenol rice cultivation

비교예 9의 분쇄 미강을 사용한 것을 제외하고는 실시예 6과 동일한 방법으로 백강균을 배양하였다.Bacillus bacteria were cultured in the same manner as in Example 6, except that the ground rice bran of Comparative Example 9 was used.

비교예 14: 분쇄Comparative Example 14: Crushing + 멸균 + 폴리페놀 추가 미강 배양+ Sterilization + Add polyphenol rice cultivation

비교예 9의 분쇄 미강을 사용한 것을 제외하고는 실시예 7과 동일한 방법으로 백강균을 배양하였다.Bacillus bacteria were cultured in the same manner as in Example 7, except that the ground rice bran of Comparative Example 9 was used.

실시예 1 내지 7, 및 비교예 1 내지 14의 백강균 배양조건을 아래의 표 1에 정리하였다.Examples 1 to 7, and Comparative Examples 1 to 14 are summarized in Table 1 below for the culture conditions of Bacillus bacteria.

구분division 배지 재료Badge material 추가분쇄여부Whether additional grinding 멸균처리여부Sterilization 폴리페놀추가여부Whether polyphenol is added 실시예 1Example 1 커피박Coffee leaf ×× ×× ×× 실시예 2Example 2 ○○ ×× ×× 실시예 3Example 3 ×× ○○ ×× 실시예 4Example 4 ○○ ○○ ×× 실시예 5Example 5 ×× ○○ ○○ 실시예 6Example 6 ○○ ×× ○○ 실시예 7Example 7 ○○ ○○ ○○ 비교예 1Comparative Example 1 아몬드박Almond gourd ×× ×× ×× 비교예 2Comparative Example 2 ○○ ×× ×× 비교예 3Comparative Example 3 ×× ○○ ×× 비교예 4Comparative Example 4 ○○ ○○ ×× 비교예 5Comparative Example 5 ×× ○○ ○○ 비교예 6Comparative Example 6 ○○ ×× ○○ 비교예 7Comparative Example 7 ○○ ○○ ○○ 비교예 8Comparative Example 8 미강Rice bran ×× ×× ×× 비교예 9Comparative Example 9 ○○ ×× ×× 비교예 10Comparative Example 10 ×× ○○ ×× 비교예 11Comparative Example 11 ○○ ○○ ×× 비교예 12Comparative Example 12 ×× ○○ ○○ 비교예 13Comparative Example 13 ○○ ×× ○○ 비교예 14Comparative Example 14 ○○ ○○ ○○

[실험예][Experimental Example]

실험예 1: 배지 재료의 분쇄에 따른 배양 효율 분석Experimental Example 1: Analysis of cultivation efficiency according to pulverization of medium material

백강균 배양용 배지 제조시 커피박, 아몬드박 또는 미강에 대하여 추가 분쇄처리를 한 것을 사용한 경우와 분쇄처리를 하지 않은 경우 최종 배양된 백강균의 균주 농도를 비교하여 아래의 표 2에 나타내었다.When preparing a culture medium for Bacillus cultivation, coffee meal, almond meal, or rice bran were subjected to additional pulverization treatment and the concentration of the final cultured Bacillus bacteria was compared and shown in Table 2 below.

구분division 배지 재료Badge material 추가분쇄여부Whether additional grinding 멸균처리여부Sterilization 폴리페놀추가여부Whether polyphenol is added 균주 농도(CFU/㎖)Strain concentration (CFU/ml) 실시예 5Example 5 커피박Coffee leaf ×× ○○ ○○ 1.02 × 10 9 1.02 × 10 9 실시예 7Example 7 ○○ ○○ ○○ 1.07 × 10 9 1.07 × 10 9 비교예 5Comparative Example 5 아몬드박Almond gourd ×× ○○ ○○ 2.88 × 10 7 2.88 × 10 7 비교예 7Comparative Example 7 ○○ ○○ ○○ 1.20 × 10 8 1.20 × 10 8 비교예 12Comparative Example 12 미강Rice bran ×× ○○ ○○ 3.72 × 10 7 3.72 × 10 7 비교예 14Comparative Example 14 ○○ ○○ ○○ 1.26 × 10 8 1.26 × 10 8

먼저, 배지 재료에 따른 배양 효율을 비교하면, 미분쇄 커피박을 사용한 실시예 5는 동일 조건의 아몬드박을 사용한 비교예 5에 비해 12배 이상, 미강을 사용한 비교예 12에 비하여 15배 이상 최종 균주 농도가 높은 것으로 측정되었다. 한편, 각각의 재료에서 추가 분쇄 여부에 따른 배양 효율을 비교하면, 커피박을 사용하는 실시예 5와 실시예 7은 분쇄 여부와 균주 농도는 유의적인 차이를 나타내지 않았다. 즉, 커피박을 배지 재료로 사용하는 경우에는 추가 분쇄 공정이 필요하지 않음을 알 수 있었다. 또한, 아몬드박을 사용하는 비교예 5와 비교예 7을 비교하면, 분쇄 아몬드박을 사용한 비교예 7이 미분쇄 아몬드박을 사용한 비교예 5에 비하여 최종 균주 농도가 4배 이상 높은 것으로 나타났다. 즉, 아몬드박을 배지 재료로 사용하는 경우에는 추가 분쇄 공정을 수행함으로써 4배 이상의 배양 효율을 나타낼 수 있음을 의미하고, 아몬드박은 분쇄 공정을 수행하여야만 균주 생산량을 늘릴 수 있다는 것을 알 수 있었다. 또한, 미강을 사용한 비교예 12와 비교예 14를 비교하면, 분쇄 미강을 사용한 비교예 14가 미분쇄 미강을 사용한 비교예 12에 비하여 4배 이상의 최종 균주 농도를 나타내었다. 미강 또한 추가 분쇄 공정을 수행하여야만 균주 생산량을 늘릴 수 있음을 알 수 있었다.First, when comparing the cultivation efficiency according to the medium material, Example 5 using pulverized coffee foil was 12 times or more compared to Comparative Example 5 using almond meal under the same conditions, and 15 times or more compared to Comparative Example 12 using rice bran It was determined that the strain concentration was high. On the other hand, when comparing the cultivation efficiency according to whether or not further pulverization in each material, Examples 5 and 7 using coffee grounds did not show a significant difference in pulverization status and strain concentration. In other words, it was found that when the coffee leaf was used as a medium material, an additional grinding process was not required. In addition, when comparing Comparative Example 5 and Comparative Example 7 using almond meal, it was found that Comparative Example 7 using crushed almond meal had a final strain concentration four or more times higher than that of Comparative Example 5 using pulverized almond meal. That is, in the case of using almond meal as a medium material, it was found that the cultivation efficiency of four or more times could be exhibited by performing an additional pulverization process, and it was found that the production of strains could be increased only when the almond meal was pulverized. In addition, when comparing Comparative Example 12 and Comparative Example 14 using rice bran, Comparative Example 14 using pulverized rice bran showed a final strain concentration of 4 times or more compared to Comparative Example 12 using pulverized rice bran. It was found that rice bran can also increase the strain production only by performing an additional grinding process.

상기 결과로 볼 때, 본원발명의 커피박을 포함하는 배지는 다른 폐기물 재료인 아몬드박이나 미강을 포함하는 배지를 사용한 경우보다 현저히 높은 배양 효율을 나타낼 뿐 아니라, 커피박이 미세 다공 구조로 되어 있어 아몬드박이나 미강에 비해 비표면적이 높으므로 추가 분쇄 공정이 필요없어 공정 단계를 줄일 수 있는 효과가 있음을 알 수 있다. From the above results, the medium containing the coffee meal of the present invention not only exhibits significantly higher cultivation efficiency than the case of using a medium containing almond meal or rice bran, which are other waste materials, and the coffee meal has a microporous structure. It can be seen that since the specific surface area is higher than that of gourd or rice bran, there is no need for an additional pulverization process, which can reduce process steps.

실험예 2: 배지 재료의 멸균 처리에 따른 배양 효율 분석Experimental Example 2: Analysis of culture efficiency according to sterilization of medium material

백강균 배양용 배지 제조시 커피박, 아몬드박 또는 미강에 대하여 멸균 처리를 한 것과 하지 않은 경우 최종 배양된 백강균의 균주 농도를 비교하여 아래의 표 3에 나타내었다. 이때, 추가 분쇄는 아몬드박과 미강에 있어서 배양 효율을 크게 좌우하므로 일률적으로 수행한 것으로 하였고, 폴리페놀 추가는 하지 않은 것을 비교하였다.When preparing a culture medium for Bacillus bacteria culture, the concentrations of the strains of the final cultured Bacillus bacteria were compared with those obtained by sterilizing coffee meal, almond meal, or rice bran, and are shown in Table 3. At this time, since the additional pulverization greatly influenced the cultivation efficiency in the almond meal and rice bran, it was considered to be performed uniformly, and the polyphenol was not added.

구분division 배지 재료Badge material 추가분쇄여부Whether additional grinding 멸균처리여부Sterilization 폴리페놀추가여부Whether polyphenol is added 균주 농도(CFU/㎖)Strain concentration (CFU/ml) 실시예 2Example 2 커피박Coffee leaf ○○ ×× ×× 1.03 × 10 9 1.03 × 10 9 실시예 4Example 4 ○○ ○○ ×× 1.09 × 10 9 1.09 × 10 9 비교예 2Comparative Example 2 아몬드박Almond gourd ○○ ×× ×× 3.80 × 10 7 3.80 × 10 7 비교예 4Comparative Example 4 ○○ ○○ ×× 1.15 × 10 8 1.15 × 10 8 비교예 9Comparative Example 9 미강Rice bran ○○ ×× ×× 1.26 × 10 8 1.26 × 10 8 비교예 11Comparative Example 11 ○○ ○○ ×× 1.13 × 10 8 1.13 × 10 8

먼저, 배지 재료에 따른 배양 효율을 비교하면, 미분쇄 커피박을 사용한 실시예 2는 동일 조건의 아몬드박을 사용한 비교예 2에 비해 17배 이상, 미강을 사용한 비교예 9에 비하여 8배 이상 최종 균주 농도가 높은 것으로 측정되었다. 한편, 각각의 재료에서 멸균처리 여부에 따른 배양 효율을 비교하면, 커피박을 사용하는 실시예 2와 실시예 4는 멸균처리 여부와 균주 농도는 유의적인 차이를 나타내지 않았다. 즉, 커피박을 배지 재료로 사용하는 경우에는 멸균처리 공정이 필요하지 않음을 알 수 있었다. 또한, 아몬드박을 사용하는 비교예 2와 비교예 4를 비교하면, 멸균처리를 한 아몬드박을 사용한 비교예 2가 멸균처리 하지 않은 아몬드박을 사용한 비교예 4에 비하여 최종 균주 농도가 2배 가량 높은 것으로 나타났다. 즉, 아몬드박을 배지 재료로 사용하는 경우에는 멸균처리를 수행함으로써 2배 이상의 배양 효율을 나타낼 수 있음을 의미하고, 아몬드박은 멸균처리를 하여야만 균주 생산량을 늘릴 수 있다는 것을 알 수 있었다. 또한, 미강을 사용한 비교예 9와 비교예 11을 비교하면, 최종 균주 농도에 유의적인 차이가 나타나지 않았다. 미강은 아몬드박과는 달리 멸균처리가 필요하지 않았다.First, when comparing the cultivation efficiency according to the medium material, Example 2 using pulverized coffee foil was 17 times or more compared to Comparative Example 2 using almond meal under the same conditions, and 8 times or more compared to Comparative Example 9 using rice bran It was determined that the strain concentration was high. On the other hand, when comparing the cultivation efficiency according to whether or not sterilized in each material, Examples 2 and 4 using coffee foil did not show a significant difference between whether or not sterilized and the strain concentration. In other words, it was found that the sterilization treatment process was not required when coffee leaf was used as a medium material. In addition, when comparing Comparative Example 2 and Comparative Example 4 using almond meal, Comparative Example 2 using sterilized almond meal had twice the final strain concentration as compared to Comparative Example 4 using almond meal that was not sterilized. Appeared to be high. That is, in the case of using almond meal as a medium material, it was found that the cultivation efficiency of more than two times could be exhibited by performing sterilization treatment, and it was found that the production of strains could be increased only when almond meal was sterilized. In addition, when comparing Comparative Example 9 and Comparative Example 11 using rice bran, there was no significant difference in the final strain concentration. Rice bran, unlike almond meal, did not require sterilization.

상기 결과로 볼 때, 본원발명의 커피박을 포함하는 배지는 폐기물 재료인 아몬드박이나 미강을 포함하는 배지를 사용한 경우보다 현저히 높은 배양 효율을 나타낼 뿐 아니라, 커피박에는 폴리페놀이 다량 함유되어 멸균처리가 필요없어 공정 단계를 줄일 수 있는 효과가 있음을 알 수 있다. From the above results, the medium containing the coffee meal of the present invention not only exhibits significantly higher cultivation efficiency than the case of using a medium containing almond meal or rice bran, which is a waste material, but also sterilizes because a large amount of polyphenol is contained in the coffee leaf. It can be seen that there is no need for treatment, which has the effect of reducing process steps.

실험예 3: 폴리페놀 추가에 따른 배양 효율 분석Experimental Example 3: Analysis of culture efficiency according to polyphenol addition

백강균 배양용 배지 제조시 커피박, 아몬드박 또는 미강에 대하여 폴리페놀을 추가하거나 추가하지 않은 경우 최종 배양된 백강균의 균주 농도를 비교하여 아래의 표 4에 나타내었다. 이때, 상술한 바와 같이 멸균처리가 아몬드박에 있어서 배양 효율을 크게 좌우하므로 멸균처리를 한 것과 하지 않은 것에 대해 각각 폴리페놀 추가에 따른 효율을 분석하였다.When preparing a culture medium for Bacillus bacteria culture, when polyphenol was added or not added to coffee meal, almond meal, or rice bran, the concentrations of the strains of the final cultured Bacillus bacteria were compared and shown in Table 4 below. At this time, as described above, since the sterilization treatment greatly influences the cultivation efficiency in the almond meal, the efficiency according to the addition of polyphenols was analyzed for the sterilization treatment and the non-sterilization treatment.

구분division 배지 재료Badge material 추가분쇄여부Whether additional grinding 멸균처리여부Sterilization 폴리페놀추가여부Whether polyphenol is added 균주 농도(CFU/㎖)Strain concentration (CFU/ml) 실시예 2Example 2 커피박Coffee leaf ○○ ×× ×× 1.03 × 10 9 1.03 × 10 9 실시예 4Example 4 ○○ ○○ ×× 1.09 × 10 9 1.09 × 10 9 실시예 6Example 6 ○○ ×× ○○ 1.12 × 10 9 1.12 × 10 9 실시예 7Example 7 ○○ ○○ ○○ 1.07 × 10 9 1.07 × 10 9 비교예 2Comparative Example 2 아몬드박Almond gourd ○○ ×× ×× 3.80 × 10 7 3.80 × 10 7 비교예 4Comparative Example 4 ○○ ○○ ×× 1.15 × 10 8 1.15 × 10 8 비교예 6Comparative Example 6 ○○ ×× ○○ 8.22 × 10 7 8.22 × 10 7 비교예 7Comparative Example 7 ○○ ○○ ○○ 1.20 × 10 8 1.20 × 10 8 비교예 9Comparative Example 9 미강Rice bran ○○ ×× ×× 1.26 × 10 8 1.26 × 10 8 비교예 11Comparative Example 11 ○○ ○○ ×× 1.13 × 10 8 1.13 × 10 8 비교예 13Comparative Example 13 ○○ ×× ○○ 1.03 × 10 8 1.03 × 10 8 비교예 14Comparative Example 14 ○○ ○○ ○○ 1.26 × 10 8 1.26 × 10 8

먼저, 배지 재료에 따른 배양 효율을 비교하면, 폴리페놀 추가 여부에 관계없이 본원발명의 실시예 2, 4, 6 및 7은 모두 동일 조건의 아몬드박 또는 미강을 사용한 비교예들에 비하여 10배 내지 30배로 배지 재료의 배양 효율이 현저히 높게 나타났다. 한편, 각각의 재료에서 폴리페놀 추가 여부에 따른 배양 효율을 비교하면, 커피박을 사용하는 실시예 2, 4, 6 및 7에서 균주 농도는 유의적인 차이를 나타내지 않았다. 즉, 커피박을 배지 재료로 사용하는 경우에는 폴리페놀 추가가 균주 농도에 특별한 영향을 미치지 않음을 알 수 있다. 또한, 아몬드박을 사용하고 멸균처리를 하지 않은 비교예 2와 비교예 6을 비교하면, 폴리페놀을 추가한 비교예 6이 폴리페놀을 추가하지 않은 비교예 2에 비하여 2배 가량 균주 농도가 높게 측정되었다. 또한, 아몬드박을 사용하고 멸균처리를 한 비교예 4와 비교예 7을 비교하면, 폴리페놀을 추가한 비교예 7와 폴리페놀을 추가하지 않은 비교예 4에서 균주 농도의 유의적인 차이가 없었으며, 이는 멸균처리가 수행되어 폴리페놀의 효과와 중복되어 나타난 현상으로 파악된다. 또한, 미강을 사용하고 멸균처리를 하지 않은 비교예 9와 비교예 13, 멸균처리를 하지 않은 비교예 11과 비교예 14는 균주 농도에 유의적인 차이가 없었다. First, when comparing the cultivation efficiency according to the medium material, Examples 2, 4, 6 and 7 of the present invention, regardless of whether or not polyphenol was added, are 10 times to 10 times compared to Comparative Examples using almond meal or rice bran under the same conditions. The culture efficiency of the medium material was remarkably high by 30 times. On the other hand, when comparing the cultivation efficiency according to whether or not polyphenol was added in each material, the strain concentration in Examples 2, 4, 6, and 7 using coffee ground did not show a significant difference. That is, it can be seen that in the case of using coffee leaf as a medium material, addition of polyphenol does not have a special effect on the strain concentration. In addition, when comparing Comparative Example 2 and Comparative Example 6 using almond meal and not sterilized, the strain concentration of Comparative Example 6 to which polyphenol was added was twice as high as that of Comparative Example 2 to which polyphenol was not added. Was measured. In addition, when comparing Comparative Example 4 and Comparative Example 7 in which almond meal was used and sterilized, there was no significant difference in strain concentration in Comparative Example 7 in which polyphenol was added and Comparative Example 4 in which polyphenol was not added. , This is considered to be a phenomenon that appeared in duplicate with the effect of polyphenols due to sterilization treatment. In addition, there was no significant difference in strain concentration between Comparative Example 9 and Comparative Example 13 without sterilization treatment using rice bran and Comparative Example 11 and Comparative Example 14 without sterilization treatment.

상기 결과로 볼 때, 본원발명의 커피박을 포함하는 배지는 폐기물 재료인 아몬드박이나 미강을 포함하는 배지를 사용한 경우보다 현저히 높은 배양 효율을 나타낼 뿐 아니라, 커피박에는 폴리페놀이 다량 함유되어 멸균처리가 필요없어 공정 단계를 줄일 수 있는 효과가 있음을 알 수 있다. From the above results, the medium containing the coffee meal of the present invention not only exhibits significantly higher cultivation efficiency than the case of using a medium containing almond meal or rice bran, which is a waste material, but also sterilizes because a large amount of polyphenol is contained in the coffee leaf. It can be seen that there is no need for treatment, which has the effect of reducing process steps.

실험예 4: 분쇄 + 멸균 + 폴리페놀 추가 수행하지 않은 경우 배양 효율 비교Experimental Example 4: Comparison of cultivation efficiency when pulverization + sterilization + polyphenol was not added

커피박, 아몬드박 및 미강 각각에 대해 분쇄, 멸균, 및 폴리페놀 추가를 수행하지 않은 실시예 1, 비교예 1 및 비교예 8의 백강균 배양 효율을 비교하여 아래의 표 5에 나타내었다.The cultivation efficiency of Bacillus bacteria in Examples 1, Comparative Example 1 and Comparative Example 8 in which pulverization, sterilization, and polyphenol addition were not performed for each coffee meal, almond meal, and rice bran were compared and shown in Table 5 below.

구분division 배지 재료Badge material 추가분쇄여부Whether additional grinding 멸균처리여부Sterilization 폴리페놀추가여부Whether polyphenol is added 균주 농도(CFU/㎖)Strain concentration (CFU/ml) 실시예 1Example 1 커피박Coffee leaf ×× ×× ×× 1.13 × 10 9 1.13 × 10 9 비교예 1Comparative Example 1 아몬드박Almond gourd ×× ×× ×× 2.22 × 10 7 2.22 × 10 7 비교예 8Comparative Example 8 미강Rice bran ×× ×× ×× 3.45 × 10 7 3.45 × 10 7

이에 따르면, 커피박을 사용한 실시예 1의 최종 균주 농도가 아몬드박을 사용한 비교예 1과 비교하여 약 30배, 미강을 사용한 비교예 8에 비하여 약 100배의 균주 농도를 나타내었다. 다시 말해, 본원발명의 커피박을 포함하는 백강균 배양용 배지는 커피박을 주재료로 사용함으로써 분쇄나 멸균 등 추가 공정, 멸균을 위한 성분 추가 없이도 현저히 높은 백강균 배양 효율을 나타낼 수 있음을 알 수 있었다.According to this, the final strain concentration of Example 1 using coffee meal was about 30 times as compared to Comparative Example 1 using almond meal, and about 100 times as compared to Comparative Example 8 using rice bran. In other words, it was found that the medium for cultivation of Bacillus bacteria containing coffee foil of the present invention can exhibit remarkably high cultivation efficiency of Bacillus bacteria without additional processes such as pulverization or sterilization, or addition of ingredients for sterilization by using coffee leaf as a main material.

본 발명의 커피박을 포함하는 백강균 배양용 배지는 커피박을 주성분으로 하고, 커피박은 미세 기공이 매우 발달한 다공성 재료이며, 폴리페놀 함량이 높아 항균성이 있어 세균이 번식하기 어려우므로 표면적으로 넓혀주기 위해 별도로 분쇄공정을 수행하거나, 세균 번식을 방지하기 위하여 살균처리를 할 필요가 없어 배지 제조가 용이하고, 이와 같은 백강균 배양용 배지를 이용하여 백강균을 배양할 경우 종래 사용되는 미강이나 아몬드박과 같은 재료에 비해 백강균 배양 효율을 현저히 향상시킬 수 있어 대량생산을 가능케 하여 농지에서의 곤충방제로 활용하기에 유리하다.The medium for cultivation of Bacillus bacteria containing coffee leaf of the present invention is made of coffee leaf as a main component, and coffee leaf is a porous material with very developed micropores, and it has high antibacterial properties due to high polyphenol content, so it is difficult for bacteria to propagate. It is easy to prepare a medium because there is no need to perform a separate pulverization process or sterilize to prevent bacterial growth, and when cultivating white bacterium using such a medium for culturing white bacterium, it is not necessary to use rice bran or almond meal. Compared to the material, it can significantly improve the cultivation efficiency of Bacillus bacteria, enabling mass production, which is advantageous for use as insect control in farmland.

Claims (10)

커피박을 포함하는 백강균(Beauveria bassiana) 배양용 배지. Baekgang (Beauveria bassiana ) culture medium containing coffee leaf. 제1항에 있어서, 상기 배지는, 커피박 100중량부를 기준으로, 당류 0.1 내지 0.8중량부; 효모추출분말 0.1 내지 0.8중량부; 및 미네랄 성분 0.01 내지 0.2중량부;를 포함하는 것을 특징으로 하는 백강균 배양용 배지.According to claim 1, wherein the medium, based on 100 parts by weight of coffee meal, 0.1 to 0.8 parts by weight of sugar; 0.1 to 0.8 parts by weight of yeast extract powder; And 0.01 to 0.2 parts by weight of a mineral component. 제1항에 있어서, 상기 커피박은 미분쇄 상태의 커피박 분말인 것을 특징으로 하는 백강균 배양용 배지.The medium for cultivating Bacillus bacteria according to claim 1, wherein the coffee meal is a coffee meal powder in a pulverized state. 제1항에 있어서, 상기 커피박의 평균입경은 180 내지 220㎛ 인 것을 특징으로 하는 백강균 배양용 배지. The medium for culturing Bacillus bacteria according to claim 1, wherein the average particle diameter of the coffee foil is 180 to 220 μm. 제1항에 있어서, 상기 커피박은 비표면적(BET)이 0.07 내지 0.13 m2 /g 인 것을 특징으로 하는 백강균 배양용 배지.The medium for cultivating Bacillus bacteria according to claim 1, wherein the coffee meal has a specific surface area (BET) of 0.07 to 0.13 m 2 /g. 커피박을 포함하는 배지를 이용한 백강균(Beauveria bassiana)의 배양방법.Cultivation method of Baekgang (Beauveria bassiana ) using a medium containing coffee leaf. 제1항에 있어서, 상기 커피박을 포함하는 배지를 이용한 백강균(Beauveria bassiana)의 배양방법은, (a) 로스팅된 커피 원두를 열수 추출한 후 남은 커피박을 준비하는 단계; (b) 상기 커피박을 포함하는 배지에 백강균(Beauveria bassiana)을 접종하여 배양하는 1차 배양 단계; (c) 상기 1차 배양된 배지를 제습하여 제습된 배지를 준비하는 단계; 및 (d) 상기 제습된 배지에 백강균을 추가로 배양하는 2차 배양 단계;를 포함하는 것을 특징으로 하는 백강균의 배양방법.The method of claim 1, wherein the method of culturing Baegangbacteria bassiana using a medium containing coffee leaf comprises the steps of: (a) preparing the remaining coffee leaf after hot water extraction of roasted coffee beans; (b) a primary culture step of inoculating and culturing Baekgang bacteria (Beauveria bassiana ) in the medium containing the coffee meal; (c) preparing a dehumidified medium by dehumidifying the first cultured medium; And (d) a second culture step of further culturing Bacillus bacteria in the dehumidified medium. 제7항에 있어서, 단계 (b)의 1차 배양은 상기 배지의 수분함량이 25 내지 35wt%인 상태에서 시작하는 것을 특징으로 하는 백강균의 배양방법.The method of claim 7, wherein the primary culture of step (b) is started in a state in which the water content of the medium is 25 to 35 wt%. 제7항에 있어서, 단계 (c)에서 상기 제습은 1차 배양된 상기 배지의 수분함량이 5 내지 15wt%가 되도록 수행하는 것을 특징으로 하는 백강균의 배양방법.The method of claim 7, wherein the dehumidification in step (c) is performed so that the water content of the first cultured medium is 5 to 15 wt%. 제7항에 있어서, 단계 (d)에서 상기 2차 배양 전 또는 배양 중 배지에 커피박을 추가 투입하는 것을 특징으로 하는 백강균의 배양방법.The method of claim 7, wherein in step (d), coffee meal is additionally added to the medium before or during the secondary culture.
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