绿茶茶渣发酵菌株驯化选育及菌酶协同发酵的研究  

Study on domestication and breeding of green tea dreg fermentation strains and bacteria-enzyme synergistic fermentation

作　　者：黄钦耿 黄炜乾 蒋顺进 于洪巍[3] 赵颖 唐谢芳 周绍迁 王英 HUANG Qingeng;HUANG Weiqian;JIANG Shunjin;YU Hongwei;ZHAO Ying;TANG Xiefang;ZHOU Shaoqian;WANG Ying(Guangdong Rongda Biological Co.,Ltd.,Qingyuan 511500,China;Qingyuan One Alive Institute of Biological Research Co.,Ltd.,Qingyuan 511500,China;College of Chemical and Biological Engineering,Zhejiang University,Hangzhou 310027,China;Fujian Xianyangyang Biological Technology Co.,Ltd.,Ningde 352100,China;Changtu County Animal Disease Prevention and Control Center,Changtu 112500,China)

机构地区：[1]广东容大生物股份有限公司,广东清远511500 [2]清远一生自然生物研究院有限公司,广东清远511500 [3]浙江大学化学工程与生物工程学院,杭州310027 [4]福建仙洋洋生物科技有限公司,福建宁德352100 [5]昌图县动物疫病预防控制中心,辽宁昌图112500

出　　处：《黑龙江畜牧兽医》2022年第3期111-118,123,135,共10页Heilongjiang Animal Science And veterinary Medicine

基　　金：清远市科技型中小企业技术创新项目(2020DZX012)。

摘　　要：为了开发利用地源性绿茶茶渣,制备功能性菌茶生物饲料,试验首先采用常压室温等离子体(ARTP)诱变及驯化筛选的方法进行功能性菌株的定向选育,然后通过"一罐多菌"的方式对不同功能菌株进行液体混合培养,并以菌落总数、粗蛋白、酸溶蛋白、总酸及限制性氨基酸的含量为考察指标,研究不同辅料(茶渣、麸皮、脱脂米糠、豆粕)组合对菌酶协同发酵茶渣固体的影响。结果表明:控制致死率为80%左右,用ARTP诱变能够获得耐受高浓度茶渣并且生物量高、代谢产物活性好的突变菌株,并能在50 L全自动液体发酵罐中实现不同功能突变菌株的液体混合培养,培养20小时时活菌总数最高,达到3.5×10^(10)cfu/mL,显微视野中菌体类型的比例大致为球菌∶杆菌∶酵母=10∶7∶1。在茶渣∶脱脂米糠∶豆粕=7∶2∶1时,发酵产物(以湿重计)菌落总数达到0.8×10^(10)cfu/g;粗蛋白含量为11.6%;酸溶蛋白含量为5.13%,较发酵前分别增加了128.0%,总酸含量为0.97%。说明ARTP诱变及定向驯化选育的功能型菌株可以较好地利用茶渣基质中的营养成分进行一定的代谢活动,适用于菌酶协同的方式制备菌茶生物饲料,辅料组合为茶渣∶脱脂米糠∶豆粕=7∶2∶1时发酵效果最好。In order to develop and utilize local-derived green tea dregs, and to prepare for functional bacterial tea bio-feed, the experiment first adopted ARTP mutagenesis and domestication screening methods to carry out directional breeding of functional strains, and then used the "one tank of multi-bacteria" method for liquid mixed culture of different functional strains. The total number of colonies, crude protein, acid-soluble protein, total acid, and restricted amino acids were used as the inspection indicators to study the effect of different excipient combinations(tea dregs、wheat bran、defatted rice bran and soybean meal) on bacteria-enzyme synergistic fermentation of tea dreg solids. The results showed that the lethality rate was controlled by about 80%;with ARTP mutagenesis, it was possible to obtain mutant strains that could tolerate high tea dreg concentration having high biomass and good metabolite activity;the culture of mixed mutant strains with different functions could be realized in a 50 L fully automatic liquid fermentation tank. The total number of viable bacteria in the culture for 20 hours was up to 3.5×10^(10)cfu/mL, and the ratio of the types of bacteria in the microscopic field was roughly coccus∶bacillus∶yeast=10∶7∶1. With tea dregs∶defatted rice bran∶soybean meal=7∶2∶1, the number of colonies of the fermentation product(based on wet weight) reached 0.8×10^(10)cfu/g, and the crude protein content of 11.6%, and the acid-soluble protein of 5.13%, which had an increase of 128.0%, respectively, compared with the number before fermentation;the total acid was 0.97%. The results suggested that the functional strains bred by ARTP mutagenesis and directed domestication could make good use of the nutrients in the tea dreg matrix to carry out certain metabolic activities, and were suitable for the preparation of bacterial tea bio-feed in a bacteria-enzyme synergy manner. The excipients tea dregs∶defatted rice bran∶soybean meal=7∶2∶1 had the best fermentation effect.

关 键 词：茶渣 常压室温等离子体(ARTP)诱变 驯化筛选 混合培养 菌酶协同 

分 类 号：S816.6[农业科学—饲料科学] Q939.9[农业科学—畜牧学]