猪肠道中产蛋白酶解淀粉芽孢杆菌的筛选、鉴定及其酶学性质的初步研究

doi:10.16431/j.cnki.1671-7236.2017.08.026

Abstract

Abstract:

A Bacillus strains,MY-6 strain which secreted high amount of protease was isolated from healthy pig's intestines,and it was identified as Bacillus amyloliquefaciens by the methods of morphological observation, biochemical tests,16S rDNA sequence amplification and comparison, biolog identification. In order to get an initial understanding of the enzymatic properties of its protease, the dynamic relationship between protease activity and bacterial life cycle, effects of pH,temperature,metal ions and inhibitors on protease activity,and the protease hydrolysis of different substrates were investigated. The results showed that,the protease synthesis of MY-6 strain was synchronized with its spore formation which began in the early stage of the exponential growth phase. The optimum conditions for the protease was 40℃ and pH 7.0,Ca²⁺ and Mn²⁺ had a intensive effects on the protease activity, PMSF could completely inhibit the activity of protease, indicating that the protease was a serine protease.The content of TCA soluble peptides for protein hydrolyzate fish protein, soybean protein, corn protein and casein was 0.83,0.67,0.58 and 0.38 μmol/g,respectively. In conclusion,Bacillus amyloliquefaciens MY-6 strain could be used as an initial selection of probiotic Bacillus to improve protein digestion.

Key words: pig; Bacillus amyloliquefaciens; protease; enzymatic properties

CLC Number:

S828

NING Yu-chang, ZHANG Xiao-jing, QIAO Hong-xing. Screening and Identification of Protease-producing Bacillus amyloliquefaciens from Pig's Intestines and the Preliminary Study of Its Enzymatic Properties[J]. , 2017, 44(8): 2398-2407.

References

[1] 饶辉.单胃动物和反刍动物对三大营养物质的消化机理及研究热点[J].湖南饲料,2008,6:24-26.
[2] 刘秀玲,周娟,卢静义,等.断奶仔猪非病原性腹泻的原因及防治措施[J].河南畜牧兽医(综合版),2005,26(8):26-27.
[3] 张占岭,历志英.浅谈提高猪饲料利用率的几种方法[J]. 畜禽业,2012,5:31-32.
[4] 栾素军,邢焕,孙永波,等.解淀粉芽孢杆菌在畜禽养殖中的应用研究进展[J].中国畜牧兽医,2016,43(10):2615-2620.
[5] 张娟,杨彩梅,曹广添,等.解淀粉芽孢杆菌及其作为益生菌的应用[J].动物营养学报,2014,26(4):863-867.
[6] 杨桥,韩文菊,张文俊,等.一株产新型大环内酯抗生素海洋解淀粉芽孢杆菌的分离鉴定[J].中华航海医学与高气压医学杂志,2009,16(2):92-95.
[7] 计坚.解淀粉芽孢杆菌SC06介导肠道免疫机理的研究[D].杭州:浙江大学, 2011.
[8] 沈勇涛.芽孢杆菌对母猪仔猪微生态的影响及其生长条件优化[D].武汉:华中农业大学,2011.
[9] 谢为天,李红胜,刘颖,等.一株源于红树林土壤芽孢杆菌的鉴定及其蛋白酶学性质研究[J].广东农业科学, 2014,41(9):155-158.
[10] 付文娟,杨亚晋,王庆,等.高产蛋白酶菌株的筛选及其提高发酵豆粕肽含量效果研究[J].中国畜牧杂志,2013,49(23):41-46.
[11] 刘慧.现代食品微生物学实验技术[M].北京:中国轻工业出版社,2006.
[12] Xie L,Xiao Y,Gao X. Purification and characterization of a halotolerant alkaline serine protease from yl-1 isolated from traditional Chinese fish sauce[J].Food Biotechnology,2016,30(2):137-153.
[13] 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.
[14] Palumbo J D,O'Keeffe T L,Abbas H K. Isolation of maize soil and rhizosphere bacteria with antagonistic activity against Aspergillus flavus and Fusarium verticillioides[J].Journal of Food Protection,2007,70(7):1615.
[15] 王德培,孟慧,管叙龙,等.解淀粉芽孢杆菌BI2的鉴定及其对黄曲霉的抑制作用[J].天津科技大学学报, 2010,25(6):5-9.
[16] 付玉梅,许锦珍,廖群,等.Lowry法测定寡肽的研究[J]. 药物分析杂志,2011,31(4):739-741.
[17] 张凯,方热军.动物微生态制剂在无污染畜牧业上的研究与应用[J].江西饲料,2011,2:22-24.
[18] 华洵璐,钱仁界,张宪中,等.芽孢杆菌复合制剂对断奶仔猪生长、腹泻和饲料消化率的影响[J].安徽农业科学,2015,43(3):166-170.
[19] Priest F G. Extracellular enzyme synthesis in the genus Bacillus[J]. Bacteriological Reviews,1977, 41(41):711-753.
[20] 钟泓波,郇惠杰, 雷芬芬,等.产蛋白酶深海细菌的筛选及其蛋白酶酶学性质[J].食品与发酵工业,2013,39(8):108-112.
[21] 史劲松.耐冷菌Bacillus cereus SYP A2-3产冷适蛋白酶的研究[D].无锡:江南大学,2006.
[22] 李红胜,谢为天,徐春厚. 一株解淀粉芽孢杆菌的鉴定及其蛋白酶酶学性质研究[A].生态环境与畜牧业可持续发展学术研讨会暨中国畜牧兽医学会2012年学术年会和第七届全国畜牧兽医青年科技工作者学术研讨会会议论文集——T06饲料卫生与安全专题[C].2012.
[23] 怀宝东,王颖, 张东杰.枯草芽孢杆菌耐热丝氨酸蛋白酶基因的克隆与表达[J].中国生物制品学杂志, 2013,26(10):1388-1391.
[24] 高岩,王景会,李玉秋,等.枯草芽孢杆菌凝乳酶的酶学性质[J]. 吉林农业大学学报,2012,34(4):385-390.
[25] 王方华,刘亮,王永华,等.产蛋白酶菌株Bacillus subtilis GS-1发酵条件优化及蛋白酶水解应用初步研究[J].中山大学学报(自然科学版),2012,51(4):79-86.

Screening and Identification of Protease-producing Bacillus amyloliquefaciens from Pig's Intestines and the Preliminary Study of Its Enzymatic Properties

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	WEI Mingbang, BIANBA Qiongda, XIAO Qingqing, DUAN Mengqi, CHAMBA Yangzom, SHANG Peng. Cloning,Bioinformatics and Expression Analysis of CDKN1B Gene in Tibetan Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2196-2206.
[2]	ZHANG Fangwei, ZHANG Qi, LEI Liangliang, SUN Wusheng, ZHANG Di, ZHANG Yunpeng, ZHANG Jingbo, WANG Xiuquan, ZHANG Jing, ZHANG Shumin. Polymorphism of HBEGF Gene and Its Association with Reproductive Traits in Songliao Black Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2370-2379.
[3]	PAN Pengcheng, LEI Zongquan, LI Xian, HU Xiangyun, QIN Qiantao, QIN Zhaoxian, GUAN Zhihui, CHEN Baojian, XIE Bingkun. Bioinformatics Analysis,Eukaryotic Expression Vector Construction and Tissue Expression of MYL2 Gene in Luchuan Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1796-1806.
[4]	OUYANG Kun, WEI Limin, SUN Ruiping, XIE Yali, QI Qi, LI Xiang, LIU Quanwei, CHAO Zhe, ZHANG Yongliang, CHEN Ting. Effects of Curcumin on Growth Performance,Feed Apparent Digestibility and Serum Biochemical Indexes in Hainan Tunchang Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1867-1875.
[5]	WU Jingxuan, SUN Zheng, MA Xuelian, ZHANG Liyang, LIAO Xiudong, LUO Xugang, LI Sufen, LYU Lin. A Survey on Magnesium Contents in Feed Resources for Livestock and Poultry in China [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1888-1898.
[6]	WANG Zeping, WANG Feifei, ZHANG Ruiming, FU Yanfeng, ZHAO Weimin, DAI Chaohui, CHENG Jinhua, LIAO Chao, LI Hui. Analysis on Polymorphism of SLA-1 Gene and Its Disease Resistance Potential in Suzi Pig [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1918-1927.
[7]	LIU Hongrun, ZHU Siran, FENG Lingli, ZHANG Kun, YAN Gang, WANG Yubin, ZHANG Shuai, JIANG Shan, XU Di, LAN Ganqiu, LIANG Jing. Cloning,Bioinformatics Analysis and Tissue Expression Localization of CDO1 Gene in Large White Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1352-1363.
[8]	YAO Kang, CAO Shuting, JIANG Zongyong, WANG Li. The Research Progress of Lysine Application and Requirement in Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1364-1374.
[9]	ZHANG Rui, MU Chunyu, TANG Qingping, BU Zhu, YANG Mingjun, XIA Aiping, YANG Xiaoming. Polymorphism of GnIH Gene and Its Correlation with Egg-laying Traits in Pigeons [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1412-1421.
[10]	ZHANG Fangwei, ZHANG Qi, ZHANG Yunpeng, LI Xin, ZHANG Jingbo, ZHANG Shumin, YU Yongsheng. Polymorphism of FOXA2 Gene Exon 3 and Its Association with Reproductive Traits in Songliao Black Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1444-1451.
[11]	TAN Yuancheng, LUO Hualun, ZHANG Yiyu, YANG Suan, WANG Yingtong, WANG Chunyuan. Polymorphism of MyoG Gene and Its Association with Body Size and Meat Traits in White Feather King Pigeons [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1461-1471.
[12]	HAN Dayong, ZHOU Chunbao, NI Ligang, CHEN Zhangyan, ZHAO Jinming. Genetic Diversity of mtDNA D-loop Sequence in Sujiang Pig [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1472-1479.
[13]	YANG Zhi, YU Jiaxin, YANG Linfu, WANG Kang, LI Guanzong, DUAN Gang, YANG Fumei, LIU Zhiqi, ZHOU Ke, HE Jun, DAI Feiyan. Study on the Therapeutic Effect of Modified Sini Decoction on Piglet Diarrhea [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1707-1717.
[14]	CHANG Yitong, ZHANG Wei, PENG Yinglin, CHEN Chen. Evolutionary Analysis,Target Gene Prediction and Tissue Expression Analysis of miR-192 [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 882-892.
[15]	LIU Zhen, ZHOU Xueqin, XU Fei, LI Zhefeng, SUN Hanxue, LI Guoqin, TAO Zhengrong, GUAN Mei, ZHENG Qiang, DIAO Xinping, LU Lizhi. Effects of Coated Sodium Butyrate on Growing Performance,Slaughter Performance, Immune Organ Index and Digestive Enzyme Activity of Breeding Pigeons and Squabs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 998-1006.