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滇金丝猴粪便微生物来源的β-半乳糖苷酶基因的表达及酶学性质 被引量:4
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作者 张文洪 杨雁霞 +8 位作者 杨正凤 黄遵锡 李俊俊 唐湘华 杨云娟 吴倩 慕跃林 韩楠玉 许波 《微生物学报》 CAS CSCD 北大核心 2019年第8期1561-1575,共15页
[目的]对滇金丝猴粪便微生物来源的β-半乳糖苷酶进行异源表达和纯化,并研究其酶学性质。[方法]从滇金丝猴粪便微生物的宏基因组中克隆出一个β-半乳糖苷酶基因galRBM20_1,对该基因进行异源表达和酶学性质分析。构建含有T7强启动子的pEA... [目的]对滇金丝猴粪便微生物来源的β-半乳糖苷酶进行异源表达和纯化,并研究其酶学性质。[方法]从滇金丝猴粪便微生物的宏基因组中克隆出一个β-半乳糖苷酶基因galRBM20_1,对该基因进行异源表达和酶学性质分析。构建含有T7强启动子的pEASY-E2-galRBM20_1质粒,转化至大肠杆菌BL21(DE3),经IPTG诱导表达后进行酶学性质研究。[结果]滇金丝猴粪便来源的β-半乳糖苷酶(galRBM20_1)最适pH为5.0,在pH 4-7之间能保留70%及其以上的活性。最适温度为45℃,在37℃和45℃下耐受1 h,酶活不变。特别的是,该酶具有良好的NaCl稳定性,经1-5 mol/L的NaCl作用1 h后,相对酶活均能超过初始酶活:当NaCl的作用浓度为4 mol/L时,β-半乳糖苷酶相对酶活最高(146%);当NaCl的作用浓度为5 mol/L时,β-半乳糖苷酶的相对酶活仍达到135%。[结论]本研究从滇金丝猴粪便微生物的宏基因库中克隆得到β-半乳糖苷酶基因galRBM20_1,并成功在大肠杆菌BL21(DE3)表达,首次从动物胃肠道宏基因组中获得具有耐盐和转糖基产Galactooligosaccharides(GOS)性能的β-半乳糖苷酶。该酶具有良好的耐盐性,和较广的pH作用范围,使其在食品、生物技术领域和环保方面的发展具有良好的应用价值。 展开更多
关键词 滇金丝猴 粪便微生物宏基因组 Β-半乳糖苷酶 耐盐性 转糖基
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Gut Microbiome Variation Along A Lifestyle Gradient Reveals Threats Faced by Asian Elephants
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作者 Chengbo Zhang Zhenghan Lian +5 位作者 Bo Xu Qingzhong Shen Mingwei Bao zunxi huang Hongchen Jiang Wenjun Li 《Genomics, Proteomics & Bioinformatics》 SCIE CAS CSCD 2023年第1期150-163,共14页
The gut microbiome is closely related to host nutrition and health.However,the relationships between gut microorganisms and host lifestyle are not well characterized.In the absence of confounding geographic variation,... The gut microbiome is closely related to host nutrition and health.However,the relationships between gut microorganisms and host lifestyle are not well characterized.In the absence of confounding geographic variation,we defined clear patterns of variation in the gut microbiomes of Asian elephants(AEs)in the Wild Elephant Valley,Xishuangbanna,China,along a lifestyle gradient(completely captive,semicaptive,semiwild,and completely wild).A phylogenetic analysis using the 16S rRNA gene sequences highlighted that the microbial diversity decreased as the degree of captivity increased.Furthermore,the results showed that the bacterial taxon WCHB1-41_c was substantially affected by lifestyle variations.qRT-PCR analysis revealed a paucity of genes related to butyrate production in the gut microbiome of AEs with a completely wild lifestyle,which may be due to the increased unfavorable environmental factors.Overall,these results demonstrate the distinct gut microbiome characteristics among AEs with a gradient of lifestyles and provide a basis for designing strategies to improve the well-being or conservation of this important animal species. 展开更多
关键词 Asian elephant Gut microbiome Phylogenetic analysis LIFESTYLE Host health
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