产几丁质脱乙酰酶细菌的筛选及16S rRNA分析

G. Zhou, Y. He, Huai-yun Zhang
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引用次数: 4

摘要

几丁质去乙酰化酶(CDA)是一种催化几丁质中n -乙酰- d -氨基葡萄糖对乙酰胺基水解的酶。几丁质脱乙酰酶在真菌细胞壁将几丁质转化为壳聚糖。从湘江河岸和湛江某岛的土壤样品中分离到208株CDA产菌,通过显色反应鉴定出12株CDA产菌,其中真菌3株,细菌2株,放线菌4株。通过酶活性测定筛选出产CDA最高的菌株Z7。对该菌株的16S基因进行扩增和测序。16S基因序列数据存入GenBank核苷酸序列数据库。用多序列比对软件Clustal s比对16S基因序列,用MegAlign软件计算相似度。利用MEGA进行了系统发育和分子进化分析。通过与相关菌种已发表的16S基因区进行比较,构建了葡萄发生树。在系统发育树上,菌株Z7与解淀粉芽孢杆菌的总体相似性值为99%。野生菌株属解淀粉芽孢杆菌。
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Screening and 16S rRNA Analysis of the Bacteria of Producing Chitin Deacetylase
Chitin deacetylase (CDA) is an enzyme that catalyzes the hydrolysis of acetamine groups of N-acetyl-D-glucosamine in chitin. Chitin deacetylase converting chitin to chitosan in fungal cell walls. A total of 208 strains was isolated from soil samples in the riverbank of Xiangjiang River and an island of Zhanjiang. 12 CDA producing strains was founded by color reaction that including three fungus, two bacterias and four actinomycetes. The highest CDA producing strain Z7 had been screened by enzyme activity assays. 16S gene of the strain was amplified and sequenced. The 16S gene sequence data were deposited in the GenBank nucleotide sequence database. The 16S gene sequences were aligned with the multiple - sequence alignment software Clustal S. Similarity values were calculated using the software MegAlign. Phylogenetic and molecular evolutionary analyses were conducted using MEGA by the neighbor - joining algorithms. Aphylogenetic tree was constructed by comparing with the published 16S gene regions of the related bacteria species. In the phylogenetic tree the overall similarity value between strain Z7 and Bacillus amyloliquefaciens are 99%. The wild strain was classfy as Bacillus amyloliquefaciens.
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