WO2021254117A1 - 贝莱斯芽孢杆菌ky01及其在降解厨余垃圾中的应用 - Google Patents

贝莱斯芽孢杆菌ky01及其在降解厨余垃圾中的应用 Download PDF

Info

Publication number
WO2021254117A1
WO2021254117A1 PCT/CN2021/096161 CN2021096161W WO2021254117A1 WO 2021254117 A1 WO2021254117 A1 WO 2021254117A1 CN 2021096161 W CN2021096161 W CN 2021096161W WO 2021254117 A1 WO2021254117 A1 WO 2021254117A1
Authority
WO
WIPO (PCT)
Prior art keywords
bacillus
velez
fermentation
kitchen waste
culture
Prior art date
Application number
PCT/CN2021/096161
Other languages
English (en)
French (fr)
Inventor
张康东
潘盛淇
李卓蕾
Original Assignee
金华康扬环境科技有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 金华康扬环境科技有限公司 filed Critical 金华康扬环境科技有限公司
Publication of WO2021254117A1 publication Critical patent/WO2021254117A1/zh

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/07—Bacillus
    • 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
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
    • B09B—DISPOSAL OF SOLID WASTE
    • B09B3/00—Destroying solid waste or transforming solid waste into something useful or harmless
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F9/00—Fertilisers from household or town refuse
    • C05F9/04—Biological compost
    • 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
    • C12N1/20—Bacteria; Culture media therefor
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
    • Y02W30/00—Technologies for solid waste management
    • Y02W30/50—Reuse, recycling or recovery technologies
    • Y02W30/78—Recycling of wood or furniture waste

Definitions

  • the invention relates to the technical field of microorganisms, in particular to a strain of Bacillus Velez KY01 and its application in degrading kitchen waste.
  • Kitchen waste is a general term for leftover meals discarded by food and beverage units such as households, hotels, restaurants, government agencies and enterprises.
  • the daily output of food waste in my country exceeds 20,000 tons.
  • the traditional treatment methods are mainly mixed and landfilled or directly used for feeding pigs. This method can easily cause cross-infection between humans and animals.
  • African Swine Fever the feeding of pig waste from food waste has been banned. Therefore, traditional treatment methods can no longer meet the current people's requirements for health, environment and hygiene.
  • Some cities in my country have begun to collect food waste in a centralized manner, and use aerobic composting or anaerobic digestion to produce biogas.
  • Biotransformation is one of the main development directions for the treatment of kitchen waste.
  • the degrading enzyme activity of microorganisms is used to promote the degradation of organic matter in the kitchen waste.
  • amylase, protease, lipase and cellulase under suitable conditions, the cycle of biotransformation of kitchen waste can be effectively shortened, the degradation efficiency of organic matter can be improved, and the degradation rate of waste can be greatly increased. And convert the residue into high-efficiency organic fertilizer. Since different bacterial species have different degrading enzyme activities and metabolic abilities under different conditions, those skilled in the art are dedicated to finding new bacterial species with excellent degradation properties.
  • the purpose of the present invention is to provide a strain of Bacillus Velez KY01 and its application in degrading kitchen waste.
  • the strain can effectively degrade kitchen waste and effectively reduce the pollution of kitchen waste.
  • the first aspect of the present invention provides a strain of Bacillus velez KY01, which is classified and named as Bacillus velezensis Bacillus velezensis, which has been deposited in the China Type Culture Collection (CCTCC), and the depositary address is China Wuhan University, Wuhan City, Hubei province, China Center for Type Culture Collection, the deposit number is CCTCCNO: M2020150, and the deposit date is May 28, 2020. It has been identified that the Velez Bacillus KY01 can effectively degrade kitchen waste.
  • CTCC China Type Culture Collection
  • the Bacillus Velez KY01 has the following properties: After being cultured on LB solid plate medium at 37°C for 48 hours, its colony is light yellow, opaque, irregular edges, uneven surface, no wrinkles in the middle, and spores. Observe the rod-shaped bacteria.
  • the second aspect of the present invention provides a fermentation method of Bacillus Velez KY01, which includes: inoculating Bacillus Velez KY01 in a fermentation medium for fermentation culture; the fermentation medium includes glucose 25g/L , Soy peptone 15g/L, corn steep liquor 15g/L, NaCl 3g/L, MgSO 4 0.2g/L.
  • the temperature of the fermentation culture is 37°C, and the culture time is 18-32h; preferably 24h.
  • Bacillus Velez KY01 undergoes a strain activation step before being inoculated into the fermentation medium.
  • an inoculum comprising the Bacillus Velez KY01 and/or its culture broth and/or its bacterial suspension and/or its fermentation product of the present invention.
  • the bacterial agent also includes a carrier.
  • the bacterial agent includes a fermentation product of Bacillus Velez KY01 and a carrier.
  • the water content in the bacterial agent is 30-35%, and the number of viable bacteria is 1.8 ⁇ 10 8 -3 ⁇ 10 8 cfu/g.
  • the concentration of the spores of Bacillus Velez KY01 in the fermentation product is 5 ⁇ 10 9 to 8 ⁇ 10 9 cfu/mL.
  • the raw material of the carrier is selected from one or more of wood chips, bagasse powder, wheat bran, wood bran, sawdust, straw chips, reed chips, and bamboo powder.
  • the preparation method of the carrier includes: after the raw materials are mixed uniformly, heat treatment to kill harmful pathogenic bacteria.
  • the raw materials of the carrier are sawdust and bagasse powder; the preparation method of the carrier is to mix the sawdust and bagasse powder in a volume ratio of 2 to 3: 2 to 3, and then at 60 to 80°C. Heat treatment to kill harmful pathogenic bacteria; preferably, the heating time is 1 to 3 hours, more preferably 1 hour.
  • the fourth aspect of the present invention provides that the Velez Bacillus KY01 or its culture broth or its bacterial suspension or its fermentation product or the bacterial agent is capable of degrading starch and/or protein and/or fat and/or cellulose. Aspects of the application.
  • the fifth aspect of the present invention provides the application of Bacillus Velez KY01 or its culture broth or its bacterial suspension or its fermentation product or the bacterial agent in degrading kitchen waste.
  • the sixth aspect of the present invention provides the application of Velez Bacillus KY01 or its culture broth or its bacterial suspension or its fermentation product or the bacterial agent in the preparation of bio-organic fertilizer.
  • the seventh aspect of the present invention provides a method for degrading kitchen waste and/or preparing bio-organic fertilizer, comprising using Bacillus Velez KY01 or its culture broth or its bacterial suspension or its fermentation product or the bacterial agent Dispose of kitchen waste.
  • condition of the treatment is aerobic.
  • the conditions of the treatment are suitable for the aerobic fermentation of the Velez Bacillus KY01.
  • the method for degrading kitchen waste and/or preparing bio-organic fertilizer includes: (1) cultivating the Bacillus Velez KY01; (2) adsorbing the product of step (1) on a carrier; (3) absorbing The product of step (2) is mixed with the kitchen waste and provides aerobic conditions.
  • the present invention has the following advantages:
  • a strain of Bacillus velezensis KY01 that can effectively degrade kitchen waste is obtained through screening.
  • This strain can simultaneously degrade starch, protein, fat and cellulose in kitchen waste, and the prepared inoculum can effectively degrade Kitchen waste, and the degradation product is bio-organic fertilizer, so that kitchen waste can be more efficiently used as resources.
  • a soil sample was obtained from the Jinhua Shuanglong Cave Scenic Area, and then the sample was inoculated into the screening medium.
  • the preparation of the screening medium was as follows: beef extract 5.0g, protein building 10.0g, water 1000ml, pH 7.0. Prepare separation medium 100ml, add appropriate amount of sterile glass beads, access 1.0g cow feces sample, and then enrich for 3 days.
  • the enriched culture solution was connected to the separation medium by gradient dilution method.
  • the separation medium was prepared as follows: 5.0g beef extract, 10.0g protein, 1000ml water, pH 7.0, 2% agar powder, and then Pick a single colony from the separation plate for streaking purification, repeat the purification 2 to 3 times, then draw the obtained purebred colony culture, keep it in the refrigerator for later use, the purified bacterial species is Velez spore Bacillus KY01.
  • amylase The activities of amylase, protease, lipase, and cellulase were identified on the strains obtained in Example 1, as follows:
  • the bacteria samples were spotted in starch agar medium, incubated at 37°C for 48 hours, and an appropriate amount of iodine solution was added to the plate, and the size of the transparent circle was measured to calculate the enzyme activity.
  • the diameter of the transparent starch circle D is 17.6mm
  • the diameter of the colony is 6.7mm
  • the bacteria samples were spotted and planted in casein agar medium. After culturing at 37°C for 48 hours, the size of the transparent circle was measured to calculate the enzyme activity.
  • the bacteria samples were spotted in cellulose-Congo red agar medium, and after culturing at 37°C for 120 hours, the size of the transparent circle was measured to calculate the enzyme activity.
  • the diameter of the cellulose hydrolysis transparent circle D is 7.1mm
  • the colony diameter is d 2mm
  • Velez Bacillus KY01 has strong amylase, protease, lipase and cellulase activities.
  • This example provides a fermentation method of Velez Bacillus KY01.
  • Slant culture Inoculate Velez Bacillus KY01 on the LB broth agar medium slant, culture at 37°C for 24h to activate.
  • Seed liquid culture Inoculate the activated Bacillus Velez KY01 in the LB broth liquid medium, and culture it on a shaker at 37° C. and 200 rpm for 24 hours to prepare the seed liquid.
  • Spore-producing fermentation culture inoculate the seed solution of Bacillus Velez KY01 in an improved spore-producing fermentation medium (glucose 25g/L, soybean peptone 15g/L, corn steep liquor 15g/L, NaCl 3g/L , MgSO 4 0.2g/L), 37°C, 200rpm shaker shaking culture for 24h to obtain the spore fermentation broth, in which the spore concentration is 6 ⁇ 10 9 cfu/mL.
  • an improved spore-producing fermentation medium (glucose 25g/L, soybean peptone 15g/L, corn steep liquor 15g/L, NaCl 3g/L , MgSO 4 0.2g/L), 37°C, 200rpm shaker shaking culture for 24h to obtain the spore fermentation broth, in which the spore concentration is 6 ⁇ 10 9 cfu/mL.
  • This example provides a fermentation method of Bacillus Velez KY01.
  • Slant culture Inoculate Velez Bacillus KY01 on the LB broth agar medium slant, culture at 37°C for 24h to activate.
  • Seed liquid culture inoculate the activated Bacillus Velez KY01 in the LB broth liquid medium, culture at 37° C., 200 rpm shaker for 18 hours, and then prepare the seed liquid.
  • Spore-producing fermentation culture inoculate the seed solution of Bacillus Velez KY01 in an improved spore-producing fermentation medium (glucose 25g/L, soybean peptone 15g/L, corn steep liquor 15g/L, NaCl 3g/L , MgSO 4 0.2g/L), 37°C, 200 rpm shaker shaking culture for 32 hours to obtain spore fermentation broth, in which the spore concentration is 8 ⁇ 10 9 cfu/mL.
  • an improved spore-producing fermentation medium (glucose 25g/L, soybean peptone 15g/L, corn steep liquor 15g/L, NaCl 3g/L , MgSO 4 0.2g/L), 37°C, 200 rpm shaker shaking culture for 32 hours to obtain spore fermentation broth, in which the spore concentration is 8 ⁇ 10 9 cfu/mL.
  • This example provides a preparation method of Velez Bacillus KY01 inoculum.
  • the spore fermentation broth prepared in Example 3 was diluted and mixed with the carrier by spraying, stirring, and mixing, wherein the ratio of the volume (L) of the spore fermentation broth to the mass (kg) of the carrier was 3:10, and the temperature was incubated at 40°C for 1 hour, and finally The water content of the inoculum is controlled between 30 and 35%, and the total effective viable number of the inoculum is 1.8 ⁇ 10 8 to 3 ⁇ 10 8 cfu/g.
  • Example 5 Take 100kg each of the inoculum and carrier prepared in Example 5, and put them into two KOYON200L biomass waste treatment equipment (Kangyang Environmental Technology Co., Ltd.). The inoculum and carrier are used as the experimental group. Control group. After turning on the equipment, put in 50kg of kitchen waste for bacterial activation for 2 days, then put in 100kg of kitchen waste every day, feed continuously for 30 days, and calculate the weight loss rate of kitchen waste. A total of three rounds of tests were performed, and the test results are shown in Table 1.
  • weight loss rate (A-B)/C ⁇ 100%.
  • A represents the total weight of the kitchen waste and the compound bacterial agent
  • B represents the total weight of the kitchen waste and the bacterial agent after treatment
  • C represents the total weight of the input kitchen waste.
  • the biological inoculum provided by the present invention can quickly degrade kitchen waste, and the final product obtained by the degradation is high-quality organic fertilizer. While achieving source reduction, it also realizes the resource utilization of kitchen waste.

Abstract

本发明涉及一株贝莱斯芽孢杆菌KY01及其在降解厨余垃圾中的应用，该菌株的分类命名为Bacillus velezensis KY01，已保藏于中国典型培养物保藏中心，保藏编号为CCTCC NO:M 2020150。该菌株能同时降解厨余垃圾中的淀粉、蛋白质、脂肪和纤维素，制成的菌剂能有效降解厨余垃圾，且降解产物为生物有机肥，使得厨余垃圾能够更加高效地被资源化利用。

Description

贝莱斯芽孢杆菌KY01及其在降解厨余垃圾中的应用 技术领域
本发明涉及微生物技术领域，具体涉及一株贝莱斯芽孢杆菌KY01及其在降解厨余垃圾中的应用。
背景技术
厨余垃圾是家庭、宾馆、饭店及机关企事业等饮食单位抛弃的剩余饭菜的通称。我国每天厨余垃圾的产生量超过2万吨，随着餐饮业的发展，厨余垃圾的产量正在快速地增长。传统的处理方式主要是混合后填埋或直接用于喂养生猪，这种方式很容易造成人畜之间疫病的交叉感染。随着非洲猪瘟的传播爆发，厨余垃圾喂养生猪已经被明令禁止。因此传统的处理方式已经不能满足目前人们对健康、环境和卫生的要求。我国部分城市已经开始集中收集厨余垃圾，并对厨余垃圾采用好氧堆肥或厌氧消化的方式制沼气。该种处理方式无法充分回收利用厨余中富含的营养成分，会造成资源浪费，而且经济效益较差。同时，由于厨余垃圾的高含水率、高含盐率和高油脂含量，还会导致堆肥品质不良以及二次污染等问题。
生物转化是处理厨余垃圾的主要发展方向之一，利用微生物的降解酶活性，促进厨余垃圾中的有机物发生降解。通过将具有淀粉酶，蛋白酶，脂肪酶和纤维素酶的菌种置于合适的条件下，能够有效缩短厨余垃圾生物转化的周期，提高有机物降解效率，从而使垃圾的降解速率大幅度提高，并将残留物转化为高效有机肥。由于不同菌种在不同条件下降解酶活性、代谢能力不一，本领域技术人员致力于寻找新的具有优异降解性能的菌种。
发明内容
本发明的目的在于提供一株贝莱斯芽孢杆菌KY01及其在降解厨余垃圾中的应用，该菌株可有效降解厨余垃圾，有效减少厨余垃圾污染。
[根据细则26改正09.06.2021]　
为此，本发明的第一方面提供了一株贝莱斯芽孢杆菌KY01,其分类命名为贝莱斯芽孢杆菌Bacillusvelezensis,己保藏于中国典型培养物保藏中心（简称CCTCC），保藏单位地址为中国湖北省武汉市武汉大学，中国典型培养物保藏中心，保藏编号为CCTCCNO:M2020150，保藏日期为2020年05月28日。经鉴定，所述贝莱斯芽孢杆菌KY01可有效降解厨余垃圾。
所述贝莱斯芽孢杆菌KY01具有下述性质：在LB固体平板培养基37℃培养48h后，其菌落呈浅黄色，不透明，边缘不整齐，表面不光滑，中间无褶皱，产芽孢，在显微镜下观察菌体杆状。
将所述贝莱斯芽孢杆菌KY01进行DNA测序，并将测序结果与相应的数据库进行比对，经过序列测定和在http://https://blast.ncbi.nlm.nih.gov/Blast.cgi网站上进行序列比对，发现该菌株的基因序列与已登记贝莱斯芽孢杆菌(Bacillus velezensis)的序列同源性最高，达到99％。因而确定该菌株为贝莱斯芽孢杆菌。将筛选得到的菌株命名为贝莱斯芽孢杆菌Bacillus velezensis KY01。通过鉴定，该贝莱斯芽孢杆菌KY01同时具有淀粉酶，蛋白酶，脂肪酶和纤维素酶活性。
本发明的第二方面，提供所述贝莱斯芽孢杆菌KY01的发酵方法，包括：将所述贝莱斯芽孢杆菌KY01接种于发酵培养基进行发酵培养；所述发酵培养基包括葡萄糖25g/L、大豆蛋白胨15g/L、玉米浆15g/L、NaCl 3g/L，MgSO 40.2g/L。
进一步，所述发酵培养的温度为37℃，培养时间为18～32h；优选为24h。
进一步，所述贝莱斯芽孢杆菌KY01在接种于发酵培养基之前经过菌种活化步骤。
本发明的第三方面，提供一种菌剂，所述菌剂包括本发明的贝莱斯芽孢杆菌KY01和/或其培养液和/或其菌悬液和/或其发酵产物。
进一步，所述菌剂还包括载体。
在具体实施方式中，所述菌剂包括贝莱斯芽孢杆菌KY01的发酵产物和载体。
进一步，所述菌剂中水分含量为30～35％，活菌数为1.8×10 8～3×10 8cfu/g。
进一步，所述发酵产物中，贝莱斯芽孢杆菌KY01芽孢的浓度为5×10 9～8×10 9cfu/mL。
进一步，所述载体的原料选自木屑、甘蔗渣粉末、麦麸、木糠、锯末、秸秆屑、芦苇屑、竹粉末中的一种或两种以上。
进一步，所述载体的制备方法包括：将各原料混合均匀后，加热处理以杀灭有害致病菌。
在具体的实施方式中，所述载体的原料为木屑和甘蔗渣粉末；所述载体的制备方法为将木屑与甘蔗渣粉末按体积比2～3:2～3混合均匀后，60～80℃加热处理以杀灭有害致病菌；优选地，加热时间为1～3h，更优选为1h。
本发明的第四方面，提供所述贝莱斯芽孢杆菌KY01或其培养液或其菌悬液或其发酵产物或所述菌剂在降解淀粉和/或蛋白质和/或脂肪和/或纤维素方面的应用。
本发明的第五方面，提供贝莱斯芽孢杆菌KY01或其培养液或其菌悬液或其发酵产物或所述菌剂在降解厨余垃圾中的应用。
本发明的第六方面，提供贝莱斯芽孢杆菌KY01或其培养液或其菌悬液或其发酵产物或所述菌剂在制备生物有机肥方面的应用。
本发明的第七方面，提供一种降解厨余垃圾和/或制备生物有机肥的方法，包括用贝莱斯芽孢杆菌KY01或其培养液或其菌悬液或其发酵产物或所述菌剂对厨余垃圾进行处理。
进一步，所述处理的条件为有氧。
进一步，所述处理的条件适于所述贝莱斯芽孢杆菌KY01进行有氧发酵。
进一步，所述降解厨余垃圾和/或制备生物有机肥的方法包括：(1)培养所述贝莱斯芽孢杆菌KY01；(2)将步骤(1)的产物吸附于载体；(3)将步骤(2)的产物与所述厨余垃圾混合并提供有氧条件。
与现有技术相比，本发明具有以下优点：
本发明经筛选得到一株能有效降解厨余垃圾的贝莱斯芽孢杆菌Bacillus velezensis KY01，该菌株能同时降解厨余垃圾中的淀粉、蛋白质、脂肪和纤维素，制成的菌剂能有效降解厨余垃圾，且降解产物为生物有机肥，使得厨余垃圾能够更加高效地被资源化利用。
具体实施方式
下面将更详细地描述本公开的示例性实施方式。应当理解，可以以各种形式实现本公开而不应被这里阐述的实施方式所限制。相反，提供这些实施方式是为了能够更透彻地理解本公开，并且能够将本公开的范围完整的传达给本领域的技术人员。
实施例1
从金华双龙洞景区获得土壤样品，然后将样品接种到筛选培养基中，筛选培养基的配制如下：牛肉膏5.0g，蛋白栋10.0g，水1000ml，pH值7.0。配制分离培养基100ml，加入适量无菌玻璃珠，接入1.0g牛粪样品，然后进行富集3天。
将富集的培养液，用梯度稀释法，接入分离培养基中，分离培养基的配制如下：牛肉膏5.0g，蛋白栋10.0g，水1000ml，pH值7.0，2％的琼脂粉，然后从分离平板中挑出单菌落进行划线纯化，反复纯化2～3次，然后将得到的纯种的菌落画线培养，放在冰箱中留存备用，纯化得的菌种即为贝莱斯芽孢杆菌KY01。
实施例2
对实施例1得到的菌种进行淀粉酶、蛋白酶、脂肪酶、纤维素酶活力的鉴定，具体如下：
(1)淀粉降解能力检测
取菌样点种于淀粉琼脂培养基中，37℃培养48h后在平板中加入适量碘液，测量透明圈大小计算酶活。
经测定，淀粉透明圈直径D 17.6mm，菌落直径d 6.7mm，酶活U＝(D/d)^2＝6.9。
(2)蛋白质降解能力检测
取菌样点种于酪蛋白琼脂培养基中，37℃培养48h后，测量透明圈大小计算酶活。
经测定，酪蛋白透明圈直径D 10.8，菌落直径d 2mm，酶活U＝(D/d)^2＝29.2。
(3脂肪降解能力检测
取菌样点种于橄榄油-中性红琼脂培养基中，37℃培养48h后，观察菌落与菌落周围培养基的颜色，是否变为红色。
经观察，菌落与菌落所处位置显红色
(4)纤维素降解能力检测
取菌样点种于纤维素-刚果红琼脂培养基中，37℃培养120h后，测量透明圈大小计算酶活。
经测定，纤维素水解透明圈直径D 7.1mm，菌落直径d 2mm，酶活 U＝(D/d)^2＝12.6
综上，经以上鉴定，贝莱斯芽孢杆菌KY01具有很强的淀粉酶，蛋白酶，脂肪酶和纤维素酶活力。
实施例3
本实施例提供了贝莱斯芽孢杆菌KY01的发酵方法。
(1)斜面培养：将贝莱斯芽孢杆菌KY01接种于LB肉汤琼脂培养基斜面，37℃培养24h活化。
(2)种子液培养：将活化好的贝莱斯芽孢杆菌KY01接种于LB肉汤液体培养基中，37℃，200rpm摇床振荡培养24h，即制备得到种子液。
(3)产芽孢发酵培养：将贝莱斯芽孢杆菌KY01的种子液分别接种于改良的产芽孢发酵培养基(葡萄糖25g/L、大豆蛋白胨15g/L、玉米浆15g/L、NaCl 3g/L，MgSO 4 0.2g/L)，37℃，200rpm摇床振荡培养24h，得到芽孢发酵液，其中芽孢浓度为6×10 9cfu/mL。
实施例4
本实施例提供了贝莱斯芽孢杆菌KY01发酵方法。
(1)斜面培养：将贝莱斯芽孢杆菌KY01接种于LB肉汤琼脂培养基斜面，37℃培养24h活化。
(2)种子液培养：将活化好的贝莱斯芽孢杆菌KY01接种于LB肉汤液体培养基中，37℃，200rpm摇床振荡培养18h，即制备得到种子液。
(3)产芽孢发酵培养：将贝莱斯芽孢杆菌KY01的种子液分别接种于改良的产芽孢发酵培养基(葡萄糖25g/L、大豆蛋白胨15g/L、玉米浆15g/L、NaCl 3g/L，MgSO 4 0.2g/L)，37℃，200rpm摇床振荡培养32h，得到芽孢发酵液，其中芽孢浓度为8×10 9cfu/mL。
实施例5
本实施例提供了贝莱斯芽孢杆菌KY01菌剂的制备方法。
将木屑与甘蔗渣粉末按体积比3比2混合均匀后，60℃加热处理1h杀灭有害致病菌，即制备得到载体，将其用于贝莱斯芽孢杆菌KY01菌剂的制备。
将实施例3制备得到的芽孢发酵液稀释后与载体进行喷雾搅拌混匀，其中芽孢发酵液的体积(L)与载体的质量(kg)之比为3:10，40℃保温处理1h， 最终将菌剂水分控制在30～35％之间，菌剂总有效活菌数1.8×10 8～3×10 8cfu/g。
实施例6
分别取实施例5制备得到的菌剂和载体各100kg，分别投放到两台KOYON200L型生物质垃圾处理设备(康扬环境科技有限公司)中，将投放菌剂的作为实验组，投放载体的作为对照组。将设备开启后，投放50kg厨余垃圾进行菌剂活化2天，之后每天投放100kg厨余垃圾，连续投料30天并计算厨余垃圾减重率。总共进行三轮测试，检测结果如表1所示。
其中，厨余垃圾降解率计算采用以下公式进行：减重率＝(A-B)/C×100％。其中A表示厨余垃圾、复合菌剂投入后的总重量；B表示厨余垃圾、菌剂处理后的总重量；C表示投入的餐厨垃圾的总重量。
表1
Figure PCTCN2021096161-appb-000001
由表1可知，实验组的平均减重率为84.3％。将实验组30天后出料送样专业检测机构检测，产物经检测认证为高品质有机肥，有机质质量分数高达97.2％，总养分为5.26％，完全符合《生物有机肥》NY884-2012标准。
综上可知，本发明提供的生物菌剂能快速降解厨余垃圾，降解得到的最终产物是高品质有机肥，在做到源头减量的同时，还实现了厨余垃圾的资源化利用。
以上所述，仅为本发明较佳的具体实施方式，但本发明的保护范围并不局限于此，任何熟悉本技术领域的技术人员在本发明揭露的技术范围内，可轻易想到的变化或替换，都应涵盖在本发明的保护范围之内。因此，本发明的保护范围应以所述权利要求的保护范围为准。

Claims (10)

  1. 贝莱斯芽孢杆菌KY01，其保藏于中国典型培养物保藏中心，且具有保藏编号CCTCC NO:M 2020150。
  2. 权利要求1所述的贝莱斯芽孢杆菌KY01的发酵方法，其特征在于，包括：将所述贝莱斯芽孢杆菌KY01接种于发酵培养基进行发酵培养；所述发酵培养基包括葡萄糖25g/L、大豆蛋白胨15g/L、玉米浆15g/L、NaCl 3g/L，MgSO 40.2g/L；
    优选地，所述发酵培养的温度为37℃，培养时间为18～32h；
    优选地，所述贝莱斯芽孢杆菌KY01在接种于发酵培养基之前经过菌种活化步骤。
  3. 一种菌剂，其特征在于，所述菌剂包括权利要求1所述的贝莱斯芽孢杆菌KY01和/或其培养液和/或其菌悬液和/或其发酵产物；
    优选地，所述菌剂还包括载体。
  4. 如权利要求3所述的菌剂，其特征在于，所述菌剂中水分含量为30～35％，活菌数为1.8×10 8～3×10 8cfu/g。
  5. 如权利要求3或4所述的菌剂，其特征在于，所述载体的原料选自木屑、甘蔗渣粉末、麦麸、木糠、锯末、秸秆屑、芦苇屑、竹粉末中的一种或两种以上；
    优选地，所述载体的制备方法包括：将各原料混合均匀后，加热处理以杀灭有害致病菌；
    优选地，所述载体的制备方法为将木屑与甘蔗渣粉末按体积比2～3:2～3混合均匀后，60～80℃加热处理以杀灭有害致病菌。
  6. 权利要求1所述的贝莱斯芽孢杆菌KY01或其培养液或其菌悬液或其发酵产物或权利要求3～5任一项所述的菌剂在降解淀粉和/或蛋白质和/或脂肪和/或纤维素方面的应用。
  7. 权利要求1所述的贝莱斯芽孢杆菌KY01或其培养液或其菌悬液或其发酵产物或权利要求3～5任一项所述的菌剂在降解厨余垃圾中的应用。
  8. 权利要求1所述的贝莱斯芽孢杆菌KY01或其培养液或其菌悬液或其发酵产物或权利要求3～5任一项所述的菌剂在制备生物有机肥方面的应用。
  9. 一种降解厨余垃圾和/或制备生物有机肥的方法，其特征在于，包括用权 利要求1所述的贝莱斯芽孢杆菌KY01或其培养液或其菌悬液或其发酵产物或权利要求3～5任一项所述的菌剂对厨余垃圾进行处理。
  10. 如权利要求9所述的降解厨余垃圾和/或制备生物有机肥的方法，其特征在于，所述处理的条件为有氧；
    优选地，所述降解厨余垃圾和/或制备生物有机肥的方法包括：(1)培养所述贝莱斯芽孢杆菌KY01；(2)将步骤(1)的产物吸附于载体；(3)将步骤(2)的产物与所述厨余垃圾混合并提供有氧条件。
PCT/CN2021/096161 2020-06-16 2021-05-26 贝莱斯芽孢杆菌ky01及其在降解厨余垃圾中的应用 WO2021254117A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202010550518.8A CN111893056B (zh) 2020-06-16 2020-06-16 贝莱斯芽孢杆菌ky01及其在降解厨余垃圾中的应用
CN202010550518.8 2020-06-16

Publications (1)

Publication Number Publication Date
WO2021254117A1 true WO2021254117A1 (zh) 2021-12-23

Family

ID=73206732

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2021/096161 WO2021254117A1 (zh) 2020-06-16 2021-05-26 贝莱斯芽孢杆菌ky01及其在降解厨余垃圾中的应用

Country Status (2)

Country Link
CN (1) CN111893056B (zh)
WO (1) WO2021254117A1 (zh)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114525227A (zh) * 2022-03-15 2022-05-24 武汉楚强生物科技有限公司 一种含贝莱斯芽孢杆菌cy30的微生物菌剂及其应用
CN114561327A (zh) * 2022-03-31 2022-05-31 山东农业大学 一种纤维素降解复合菌剂及其制备方法和应用
CN115058357A (zh) * 2022-06-08 2022-09-16 北京市科学技术研究院资源环境研究所 一种用于餐厨垃圾的生物干化的菌株、筛选方法及应用
CN115786206A (zh) * 2022-12-02 2023-03-14 森诺技术有限公司 一种自产热高温好氧菌种及其在餐厨垃圾生物处理中的应用

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111893056B (zh) * 2020-06-16 2022-06-03 金华康扬环境科技有限公司 贝莱斯芽孢杆菌ky01及其在降解厨余垃圾中的应用
CN112792092A (zh) * 2020-12-16 2021-05-14 青岛尚德生物技术有限公司 帕氏乳杆菌在制备用于降解厨余垃圾的微生物菌剂中的应用
CN112662595B (zh) * 2021-01-25 2022-12-27 深圳市家家分类环保技术开发有限公司 一种厨余垃圾微生物降解菌剂及其制备方法和应用
CN113249277B (zh) * 2021-07-05 2021-09-24 广东海洋大学 一株贝雷斯芽孢杆菌在水产品肝素提取中的应用
CN114134075B (zh) * 2021-11-22 2023-11-21 天津博菲德科技有限公司 一株高产复合酶同时高效降解霉菌毒素的贝莱斯芽孢杆菌及其应用
CN114702354A (zh) * 2021-12-15 2022-07-05 中农新科(苏州)有机循环研究院有限公司 一种高值化餐厨废弃物生物有机肥的制备方法及应用
CN114574383B (zh) * 2022-01-18 2024-02-23 金华康扬环境科技有限公司 高效厨余垃圾降解复合微生物菌剂及其制备方法与应用
CN114921356B (zh) * 2022-01-18 2024-04-02 金华康扬环境科技有限公司 家用型厨余垃圾好氧堆肥复合微生物菌剂及其制备方法

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20110119386A (ko) * 2010-04-27 2011-11-02 주식회사 진상 바실러스 벨렌첸시스 ａ-68 유래 섬유소 분해효소 유전자 및 이를 도입하여 형질전환된 에셰리키아 콜리 ａ-68 균주 및 이를 이용한 섬유소 분해효소의 생산 방법
CN109645214A (zh) * 2018-11-21 2019-04-19 江苏大学 一种微生物发酵处理餐厨垃圾的方法
CN111235050A (zh) * 2019-11-08 2020-06-05 浙江大学宁波理工学院 一种厨余垃圾降解菌株、厨余垃圾处理剂及厨余垃圾降解方法
CN111893056A (zh) * 2020-06-16 2020-11-06 金华康扬环境科技有限公司 贝莱斯芽孢杆菌ky01及其在降解厨余垃圾中的应用

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8383097B2 (en) * 2006-08-11 2013-02-26 Novozymes Biologicals, Inc Bacteria cultures and compositions comprising bacteria cultures
CN110283742B (zh) * 2019-06-17 2020-12-15 北京农业生物技术研究中心 一株广谱抗病的贝莱斯芽孢杆菌及其应用

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20110119386A (ko) * 2010-04-27 2011-11-02 주식회사 진상 바실러스 벨렌첸시스 ａ-68 유래 섬유소 분해효소 유전자 및 이를 도입하여 형질전환된 에셰리키아 콜리 ａ-68 균주 및 이를 이용한 섬유소 분해효소의 생산 방법
CN109645214A (zh) * 2018-11-21 2019-04-19 江苏大学 一种微生物发酵处理餐厨垃圾的方法
CN111235050A (zh) * 2019-11-08 2020-06-05 浙江大学宁波理工学院 一种厨余垃圾降解菌株、厨余垃圾处理剂及厨余垃圾降解方法
CN111893056A (zh) * 2020-06-16 2020-11-06 金华康扬环境科技有限公司 贝莱斯芽孢杆菌ky01及其在降解厨余垃圾中的应用

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
JIANG JIAPENG: "Quality Evaluation of Aerobic Fermentation Products of Kitchen Waste and Its Application in the Diet for Carassius Auratus", MASTER THESIS, TIANJIN POLYTECHNIC UNIVERSITY, CN, 15 January 2020 (2020-01-15), CN , XP055882129, ISSN: 1674-0246 *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114525227A (zh) * 2022-03-15 2022-05-24 武汉楚强生物科技有限公司 一种含贝莱斯芽孢杆菌cy30的微生物菌剂及其应用
CN114525227B (zh) * 2022-03-15 2023-11-21 武汉楚强生物科技有限公司 一种含贝莱斯芽孢杆菌cy30的微生物菌剂及其应用
CN114561327A (zh) * 2022-03-31 2022-05-31 山东农业大学 一种纤维素降解复合菌剂及其制备方法和应用
CN114561327B (zh) * 2022-03-31 2023-09-12 山东农业大学 一种纤维素降解复合菌剂及其制备方法和应用
CN115058357A (zh) * 2022-06-08 2022-09-16 北京市科学技术研究院资源环境研究所 一种用于餐厨垃圾的生物干化的菌株、筛选方法及应用
CN115786206A (zh) * 2022-12-02 2023-03-14 森诺技术有限公司 一种自产热高温好氧菌种及其在餐厨垃圾生物处理中的应用
CN115786206B (zh) * 2022-12-02 2023-12-19 森诺技术有限公司 一种自产热高温好氧菌种及其在餐厨垃圾生物处理中的应用

Also Published As

Publication number Publication date
CN111893056A (zh) 2020-11-06
CN111893056B (zh) 2022-06-03

Similar Documents

Publication Publication Date Title
WO2021254117A1 (zh) 贝莱斯芽孢杆菌ky01及其在降解厨余垃圾中的应用
CN106011027B (zh) 一种高效降解餐厨垃圾的生物处理剂及其制备方法
JP7223808B2 (ja) 生ゴミ高温生分解用微生物接種剤、および生ゴミ分解の方法
Shukla et al. Syntrophic microbial system for ex-situ degradation of paddy straw at low temperature under controlled and natural environment
CN106047762B (zh) 一种枯草芽孢杆菌及其在餐厨垃圾中的应用
CN106190900B (zh) 一种地衣芽孢杆菌及其在餐厨垃圾中的应用
CN111235050B (zh) 一种厨余垃圾降解菌株、厨余垃圾处理剂及厨余垃圾降解方法
KR102004089B1 (ko) 음식물쓰레기 분해 혼합균주 및 이를 이용한 음식물쓰레기 분해 방법
CN103602616B (zh) 一种解淀粉芽孢杆菌y10及其应用
CN110643544B (zh) 副干酪乳杆菌761及其应用、青贮饲料添加剂、青贮饲料
CN108795814B (zh) 一种可降解废弃羽毛的菌株、筛选方法及其应用
CN110591987B (zh) 唾液乳杆菌358及其应用、青贮饲料添加剂、青贮饲料
AU2021103470A4 (en) Bacillus subtilis bs40-4 strain and method for composting organic wastes by using the same
CN114574383B (zh) 高效厨余垃圾降解复合微生物菌剂及其制备方法与应用
CN107988099B (zh) 一种有机垃圾快速降解减量化的微生物菌剂及其应用
CN107365718B (zh) 巨大芽孢杆菌myb3及其在秸秆发酵饲料中的应用
CN113481118B (zh) 一株耐酸、耐盐、耐热、降解有机物的枯草芽孢杆菌及其在餐厨垃圾资源化中的应用
CN109402013B (zh) 一种产脂肪酶的耐高温枯草芽孢杆菌及其在堆肥中的应用
CN105950483B (zh) 一种光滑假丝酵母及其在餐厨垃圾中的应用
CN110055197B (zh) 一种解淀粉类芽孢杆菌brec-10及其菌剂和应用
CN106148233B (zh) 一种乳酸片球菌及其在餐厨垃圾中的应用
CN106085915B (zh) 一种金黄短杆菌及其在餐厨垃圾中的应用
CN111269860A (zh) 一种降解餐厨垃圾的微生物菌种及其应用
CN104560817B (zh) 一株产植酸酶的嗜热地衣芽孢杆菌utm102及其应用
CN112760274A (zh) 一种有机固废高温好氧堆肥菌种及其应用

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 21825085

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 21825085

Country of ref document: EP

Kind code of ref document: A1