Isolation and Application of Microbes Capable of Decomposing Tea Dregs

MIAO Furong; CHEN Xinzhu; LI Zhongrong; LIU Jing

doi:10.19303/j.issn.1008-0384.2021.04.015

Volume 36 Issue 4

Apr. 2021

Turn off MathJax

Article Contents

Article Navigation > Fujian Journal of Agricultural Sciences > 2021 > 36(4): 480-489

MIAO F R, CHEN X Z, LI Z R, et al. Isolation and Application of Microbes Capable of Decomposing Tea Dregs [J]. Fujian Journal of Agricultural Sciences，2021，36（4）：480−489 doi: 10.19303/j.issn.1008-0384.2021.04.015

Citation:

MIAO F R, CHEN X Z, LI Z R, et al. Isolation and Application of Microbes Capable of Decomposing Tea Dregs [J]. Fujian Journal of Agricultural Sciences，2021，36（4）：480−489 doi: 10.19303/j.issn.1008-0384.2021.04.015

Citation:

PDF( 1159 KB)

Isolation and Application of Microbes Capable of Decomposing Tea Dregs

doi: 10.19303/j.issn.1008-0384.2021.04.015

Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian　350013, China

Received Date: 2020-09-02
Rev Recd Date: 2021-02-28

Available Online: 2021-04-20

Publish Date: 2021-04-30

Abstract

Abstract

Objective Microorganisms that can efficiently decompose tea dregs at high temperature were screened for potential waste treatment application. Method A high temperature special habitat separation method was employed to isolate cellulase and protease-producing bacteria on tea dregs. Morphological, physiochemical, and growth characteristics of selected strains were examined prior to 16S rDNA cloning for sequencing. Gene sequences of the candidates, along with gyrB, were subjected to phylogenetic analysis for species identification. Decomposing capacity of the isolated strains on tea dregs was determined for final selection. Results Five thermophiles exhibiting the desired properties were isolated. Among them, Fb showed the highest enzymatic activities and was found to be a strain of Bacillus velezensis. It was further characterized with a high temperature tolerance up to 55 ℃ and the optimal culture conditions of 42-45 ℃, pH 5.0-7.0, 16 h incubation, 1.0-6.0% salinity, 30 mL 250 min⁻¹ filling volume, and 120 rpm shaker speed. After Fb inoculation and incubation for 7 d, the crude protein in the resulting tea dregs significantly increased by 14.88% (P＜0.05), the contents of 14 amino acids, except cystine, methionine, and histidine, significantly increased (P＜0.05), the total amino acids increased by 5.98%, the crude fiber decreased by 9.69%, and the neutral detergent fiber, acid detergent fiber, and lignin decreased by 10.72%, 4.47%, and 11.37%, respectively (P＜0.05). Conclusion For the first time, B. velezensis Fb was identified to be capable of efficiently decomposing tea dregs with a significantly improved nutritional profile on the substrate. It could conceivably become a bioagent for treating the waste material.
- Tea dregs,
- Bacillus velezensis,
- isolation,
- protein,
- cellulose

FullText(HTML)

References(47)

References

[1]	朱飞, 苏娣, 冉雷, 等. 黑曲霉固态发酵改善茶渣营养价值的研究 [J]. 动物营养学报, 2018, 30(10):4269−4278. doi: 10.3969/j.issn.1006-267x.2018.10.054 ZHU F, SU D, RAN L, et al. Nutritional improvement of tea residue by solid-state fermentation with Aspergillus niger [J]. Chinese Journal of Animal Nutrition, 2018, 30(10): 4269−4278.(in Chinese) doi: 10.3969/j.issn.1006-267x.2018.10.054
[2]	郑清梅, 陈昆平, 钟艳梅, 等. 4类茶叶及其茶渣主要成分的测定与分析 [J]. 广东农业科学, 2015, 42(6):14−20. doi: 10.3969/j.issn.1004-874X.2015.06.003 ZHENG Q M, CHEN K P, ZHONG Y M, et al. Determination and analysis of main components of four kinds of tea and their tea wastes [J]. Guangdong Agricultural Sciences, 2015, 42(6): 14−20.(in Chinese) doi: 10.3969/j.issn.1004-874X.2015.06.003
[3]	叶勇. 茶饲料的应用及发展前景 [J]. 中国茶叶, 2000(4):14−15. YE Y. Application and development prospects of tea feed [J]. China Tea, 2000(4): 14−15.(in Chinese)
[4]	刘红云, 梁慧玲. 茶渣用作饲料的研究 [J]. 饲料研究, 2004(9):19−20. doi: 10.3969/j.issn.1002-2813.2004.09.006 LIU H Y, LIANG H L. Research on the use of tea residue as feed [J]. Feed Research, 2004(9): 19−20.(in Chinese) doi: 10.3969/j.issn.1002-2813.2004.09.006
[5]	佐野满昭, 孙希琪. 茶添加到家畜饲料中的利用效果 [J]. 农业新技术新方法译丛, 1996, 0(2):32−33. SANO MANZHAO, SUN X Q. Utilization effect of tea added to livestock feed [J]. Translation of new agricultural technologies and methods, 1996, 0(2): 32−33.(in Chinese)
[6]	徐瑞, 王改琴, 邬本成, 等. 茶叶渣对育肥猪生产性能及猪肉品质的影响 [J]. 黑龙江畜牧兽医, 2017(10):73−75, 286. XU R, WANG G Q, WU B C, et al. Effect of tea residue on fattening pig performance and pork quality [J]. Heilongjiang Animal Science and Veterinary Medicine, 2017(10): 73−75, 286.(in Chinese)
[7]	吴慧敏. 茶渣、茶末对蛋鸡生产性能及鸡蛋品质的影响研究[D]. 福州: 福建农林大学, 2016. WU H M. Study on the effect of tea residue and the tea dust on production performance and egg quality[D]. Fuzhou: Fujian Agriculture and Forestry University, 2016. (in Chinese)
[8]	舒庆龄, 赵和涛. 茶渣饲养肉用鸡效果研究 [J]. 安徽农业科学, 1995(4):355−356. SHU Q L, ZHAO H T. Research on the effect of tea residues in breeding chicken for meat [J]. Journal of Anhui Agricultural Sciences, 1995(4): 355−356.(in Chinese)
[9]	潘发明, 李发弟, 郝正里, 等. 茶渣单宁含量对绵羊养分消化利用与氮代谢参数的影响 [J]. 畜牧兽医学报, 2012, 43(1):71−81. PAN F M, LI F D, HAO Z L, et al. Effects of tannin content in residue of tea-leaves on digestion and utilization of nutrients and metabolic parameters of nitrogen in sheep [J]. Chinese Journal of Animal and Veterinary Sciences, 2012, 43(1): 71−81.(in Chinese)
[10]	傅志民, 吴永福. 废弃茶渣综合再利用研究进展 [J]. 中国茶叶加工, 2011(1):17−20. FU Z M, WU Y F. Research on multi-purpose utilization of used waste tea leaves [J]. China Tea Processing, 2011(1): 17−20.(in Chinese)
[11]	谢月琴, 罗君谊, 陈婷, 等. 茶渣资源再利用研究概况 [J]. 中国农学通报, 2019, 35(33):141−145. doi: 10.11924/j.issn.1000-6850.casb18060083 XIE Y Q, LUO J Y, CHEN T, et al. Reuse of tea residue resources: A review [J]. Chinese Agricultural Science Bulletin, 2019, 35(33): 141−145.(in Chinese) doi: 10.11924/j.issn.1000-6850.casb18060083
[12]	刘姝, 涂国全. 茶渣经微生物固体发酵成饲料的初步研究 [J]. 江西农业大学学报, 2001(1):130−133. doi: 10.3969/j.issn.1000-2286.2001.01.029 LIU S, TU G Q. A preliminary study on the microbial feed by solid fermentation of tea residue [J]. Acta Agriculturae Universitis Jiangxiensis, 2001(1): 130−133.(in Chinese) doi: 10.3969/j.issn.1000-2286.2001.01.029
[13]	胡桂萍, 杨广, 欧阳雪灵. 茶渣发酵蛋白饲料辅料优选及成本分析 [J]. 重庆理工大学学报(自然科学), 2017, 31(5):86−90, 142. HU G P, YANG G, OUYANG X L. Optimation of supplemental material in microbial fermentation of tea residue and its costs analysis [J]. Journal of Chongqing University of Technology (Natural Science), 2017, 31(5): 86−90, 142.(in Chinese)
[14]	倪星虹. 茶渣提取蛋白及饲料化利用的初步研究[D]. 南京: 南京农业大学, 2011. NI X H. Studys of the protein extract and utilization of producing protein feed from tea residues[D]. Nanjing: Nanjing Agricultural University, 2011. (in Chinese)
[15]	方呈祥. 中国高等学校菌种目录[M]. 北京: 化学工业出版社, 2009.
[16]	JAIN D K, COLLINS-THOMPSON D L, LEE H, et al. A drop-collapsing test for screening surfactant-producing microorganisms [J]. Journal of Microbiological Methods, 1991, 13(4): 271−279. doi: 10.1016/0167-7012(91)90064-W
[17]	PINCHUK I V, BRESSOLLIER P, SOROKULOVA I B, et al. Amicoumacin antibiotic production and genetic diversity of Bacillus subtilis strains isolated from different habitats [J]. Research in Microbiology, 2002, 153(5): 269−276. doi: 10.1016/S0923-2508(02)01320-7
[18]	WANG L T, LEE F L, TAI C J, et al. Bacillus velezensis is a later heterotypic synonym of Bacillus amyloliquefaciens [J]. International Journal of Systematic and Evolutionary Microbiology, 2008, 58(Pt 3): 671−675.
[19]	王伟, 李术娜, 李红亚, 等. 番茄灰霉病拮抗细菌的筛选与X-75菌株鉴定 [J]. 园艺学报, 2010, 37(2):307−312. WANG W, LI S N, LI H Y, et al. Screening of antagonistic bacteria against Botrytis cinerea and identification of strain X-75 [J]. Acta Horticulturae Sinica, 2010, 37(2): 307−312.(in Chinese)
[20]	王青华, 孙晓晖, 唐旭, 等. 深海贝莱斯芽孢杆菌DH82的筛选、鉴定及其抗菌粗蛋白性质分析 [J]. 海洋通报, 2019, 38(1):63−69. WANG Q H, SUN X H, TANG X, et al. Screening and identification of Bacillus velezensis strain DH82 and the characterization of the crude antimicrobial protein [J]. Marine Science Bulletin, 2019, 38(1): 63−69.(in Chinese)
[21]	杜淑涛, 李术娜, 朱宝成. 白菜黑斑病拮抗细菌Bacillus velezensis DL-59的筛选鉴定及田间防效实验 [J]. 河北农业大学学报, 2010, 33(6):51−56. doi: 10.3969/j.issn.1000-1573.2010.06.011 DU S T, LI S N, ZHU B C. Screening and identification of antagonistic strain DL-59 of Bacillus velezensis against Alternaria brassicae and biocontrol efficiency [J]. Journal of Agricultural University of Hebei, 2010, 33(6): 51−56.(in Chinese) doi: 10.3969/j.issn.1000-1573.2010.06.011
[22]	李生樟, 陈颖, 杨瑞环, 等. 一株拮抗黄单胞菌的贝莱斯芽孢杆菌的分离和鉴定 [J]. 微生物学报, 2019, 59(10):1969−1983. LI S Z, CHEN Y, YANG R H, et al. Isolation and identification of a Bacillus velezensis strain against plant pathogenic Xanthomonas spp [J]. Acta Microbiologica Sinica, 2019, 59(10): 1969−1983.(in Chinese)
[23]	KANJANAMANEESATHIAN M, WIWATTANAPATAPEE R, ROTNIAM W, et al. Application of a suspension concentrate formulation of Bacillus velezensis to control root rot of hydroponicallygrown vegetables [J]. New Zealand Plant Protection, 2013, 66: 229−234. doi: 10.30843/nzpp.2013.66.5556
[24]	张彩文, 程坤, 张欣, 等. 贝莱斯芽胞杆菌(Bacillus velezensis)分类学及功能研究进展 [J]. 食品与发酵工业, 2019, 45(17):258−265. ZHANG C W, CHENG K, ZHANG X, et al. Taxonomy and functions of Bacillus velezensis: A review [J]. Food and Fermentation Industries, 2019, 45(17): 258−265.(in Chinese)
[25]	PAN H Q, LI Q L, HU J C. The complete genome sequence of Bacillus velezensis 9912D reveals its biocontrol mechanism as a novel commercial biological fungicide agent [J]. Journal of Biotechnology, 2017, 247: 25−28. doi: 10.1016/j.jbiotec.2017.02.022
[26]	KIM S Y, LEE S Y, WEON H Y, et al. Complete genome sequence of Bacillus velezensis M75, a biocontrol agent against fungal plant pathogens, isolated from cotton waste [J]. Journal of Biotechnology, 2017, 241: 112−115. doi: 10.1016/j.jbiotec.2016.11.023
[27]	王金燕, 李彬, 王印庚, 等. 刺参养殖池塘一株贝莱斯芽孢杆菌的分离及其生理特性 [J]. 中国水产科学, 2018, 25(3):567−575. doi: 10.3724/SP.J.1118.2018.17383 WANG J Y, LI B, WANG Y G, et al. Screening and characteristic analysis of Bacillus velezensis from sea cucumber(Apostichopus japonicus) ponds [J]. Journal of Fishery Sciences of China, 2018, 25(3): 567−575.(in Chinese) doi: 10.3724/SP.J.1118.2018.17383
[28]	LIU X Y, REN B, CHEN M, et al. Production and characterization of a group of bioemulsifiers from the marine Bacillus velezensis strain H3 [J]. Applied Microbiology and Biotechnology, 2010, 87(5): 1881−1893. doi: 10.1007/s00253-010-2653-9
[29]	TOMAZ C T, QUEIROZ J A. Fractionation of Trichoderma reesei cellulases by hydrophobic interaction chromatography on phenyl-sepharose [J]. Biotechnology Letters, 2004, 26(3): 223−227. doi: 10.1023/B:BILE.0000013719.92024.04
[30]	中华人民共和国国家质量监督检验检疫局, 中国国家标准化管理委员会．饲料添加剂酸性蛋白酶活力的测定分光光度法: GB/T 28715—2012 [S]．北京: 中国标准出版社, 2012．
[31]	中华人民共和国国家技术监管局．蛋白酶制剂: GB/T 23527—2009［S］．北京: 中国标准出版社, 2009.
[32]	杨健, 何永林, 刘晔华. 微生物教学中常用细菌染色方法的改进 [J]. 川北医学院学报, 2003(4):165. doi: 10.3969/j.issn.1005-3697.2003.04.150 YANG J, HE Y L, LIU Y H. Improvement of commonly used bacterial staining methods in microbiology teaching [J]. Journal of North Sichuan Medical College, 2003(4): 165.(in Chinese) doi: 10.3969/j.issn.1005-3697.2003.04.150
[33]	布坎南 R E,吉本斯 N E.伯杰细菌鉴定手册[M].8版.中国科学院微生物研究所《伯杰细菌鉴定手册》翻译组,译.北京: 科学出版社,1984.
[34]	沈萍. 微生物学实验[M]. 北京: 高等教育出版社, 2004.
[35]	东秀珠, 蔡妙英. 常见细菌系统鉴定手册[M]. 北京: 科学出版社, 2001: 364−398.
[36]	郝云婕, 韩素贞. gyrB基因在细菌系统发育分析中的应用 [J]. 生物技术通报, 2008(2):39−41. HAO Y J, HAN S Z. Application of gyrB gene in bacterial phylogenetic analysis [J]. Biotechnology Bulletin, 2008(2): 39−41.(in Chinese)
[37]	TANG X L, ZHAI L, LIN Y F, et al. Halomonas alkalicola sp. nov., isolated from a household product plant [J]. International Journal of Systematic and Evolutionary Microbiology, 2017, 67(5): 1546−1550. doi: 10.1099/ijsem.0.001757
[38]	PHENG S, LEE J J, EOM M K, et al. Paucibacter oligotrophus sp. nov., isolated from fresh water, and emended description of the genus Paucibacter [J]. International Journal of Systematic and Evolutionary Microbiology, 2017, 67(7): 2231−2235. doi: 10.1099/ijsem.0.001931
[39]	卫扬保. 微生物生理学[M]. 北京: 高等教育出版社, 1989.
[40]	王加启, 于建国. 饲料分析与检验[M]. 北京: 中国计量出版社, 2004.
[41]	于泓, 牟世芬. 氨基酸分析方法的研究进展 [J]. 分析化学, 2005(3):398−404. doi: 10.3321/j.issn:0253-3820.2005.03.028 YU H, MOU S F. Method development for amino acid analysis [J]. Chinese Journal of Analytical Chemistry, 2005(3): 398−404.(in Chinese) doi: 10.3321/j.issn:0253-3820.2005.03.028
[42]	BHATTACHARYA S S, GARLAPATI V K, BANERJEE R. Optimization of laccase production using response surface methodology coupled with differential evolution [J]. New Biotechnology, 2011, 28(1): 31−39. doi: 10.1016/j.nbt.2010.06.001
[43]	胡宝东, 邱树毅, 周鸿翔, 等. 酱香型大曲的理化指标、水解酶系、微生物产酶的关系研究 [J]. 现代食品科技, 2017, 33(2):99−106. HU B D, QIU S Y, ZHOU H X, et al. Relationships among physiochemical indices and hydrolyzing enzyme systems and enzymes-produced-ability in Jiangxiang daqu [J]. Modern Food Science and Technology, 2017, 33(2): 99−106.(in Chinese)
[44]	李芳蓉, 陈鑫, 张建军. 红芪药渣固态发酵生产蛋白饲料的探讨 [J]. 中央民族大学学报(自然科学版), 2019, 28(1):47−51. LI F R, CHEN X, ZHANG J J. Discussion on the production of protein feed by solid ferm entation of Hedysarum polybotrys hand. -Mazz. Residue [J]. Journal of Minzu University of China (Natural Sciences Edition), 2019, 28(1): 47−51.(in Chinese)
[45]	张沛, 杨国辉, 魏丽娟, 等. 稻壳粉固态发酵生产蛋白饲料的工艺研究 [J]. 中国饲料, 2015(8):25−27, 38. ZHANG P, YANG G H, WEI L J, et al. Study on the technology of protein feed frim rice husk powder fermented by solid slate [J]. China Feed, 2015(8): 25−27, 38.(in Chinese)
[46]	张楠, 张凌云. 茶渣在自然发酵与添加外源微生物发酵过程中的理化动态研究[C]//2009年中国茶叶科技创新与产业发展学术研讨会论文集. 重庆, 2009: 428-435.
[47]	陈奕业, 朱妮, 梁瑶, 等. 不同添加剂对凉茶渣发酵品质的影响 [J]. 家畜生态学报, 2020, 41(2):55−59. doi: 10.3969/j.issn.1673-1182.2020.02.010 CHEN Y Y, ZHU N, LIANG Y, et al. Effect of additives on quality of herbal tea residue fermentation [J]. Journal of Domestic Animal Ecology, 2020, 41(2): 55−59.(in Chinese) doi: 10.3969/j.issn.1673-1182.2020.02.010