产脂肪酶菌株的分离鉴定及其脂肪酶A基因(Lipase A)序列分析
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- 英文论文题名：Isolation and identification of lipase producing bacteria and analysis of lipase A gene
- 论文作者：古从伟 ; 何敏 ; 王振华 ; 潘康成 ; 冯杰 ; 唐慧琴 ; 张飞燕
- 英文论文作者：Congwei Gu ; Min He ; Zhenhua Wang ; Kangcheng Pan ; Jie Feng ; Huiqin Tang ; Feiyan Zhang ; Animal Microecology Institute ; Sichuan Agricultural University ; Southwest Medical University ; Deyang Animal Husbandry and Food Bureau ; Chengdu agricultural science and technology Career Academy
- 年：2016
- 作者机构：四川农业大学动物微生态研究中心;西南医科大学;德阳市畜牧食品局;成都农业科技职业学院;
- 论文关键词：铜绿假单胞菌 ; 筛选 ; 脂肪酶 ; 脂肪酶基因 ; 克隆
- 英文论文关键词：Pseudomonas aeruginosa ; Screening ; lipase ; Lipase gene ; Clone
- 会议召开时间：2016-11-04
- 会议录名称：中国畜牧兽医学会动物微生态学分会第五届第十二次全国学术研讨会论文集
- 语种：中文
- 分类号：Q93
- 学会代码：ZGXJ
- 会议名称：中国畜牧兽医学会动物微生态学分会第五届第十二次全国学术研讨会
- 会议地点：中国山东青岛
- 主办单位：中国畜牧兽医学会动物微生态学分会
- 学会名称：中国畜牧兽医学会
- 页数：1
- 文件大小：80k
- 原文格式：O
- 会议级别：全国
摘要
文章旨在筛选高产脂肪酶能力的细菌,并对其脂肪酶A基因进行克隆测序分析。采用含有维多利亚蓝的分离培养基平板从富油脂的土壤中分离到筛选得到相对高产的脂肪酶细菌,利用碱滴定法测定其酶活力大小,通过细菌形态、生理生化和细菌16S rRNA序列分析;设计脂肪酶基因引物并对脂肪酶LipA基因进行克隆测序分析。结果,筛选得到1株产脂肪酶能力相对较高的细菌A05,利用碱滴定法测定其酶活力大小为1.07 U/ml通过细菌形态、生理生化和16S rRNA序列鉴定该高产脂肪酶细菌为铜绿假单胞菌;根据GenBank中报道的铜绿假单胞菌脂肪酶A基因设计引物,成功扩增得到长1094bp的酶基因DNA序列,该序列与铜绿假单胞菌脂肪酶A基因序列同源性高达98%以上,该序列包含1个完整的开放阅读框,936bp,共编码311个氨基酸,前26个氨基酸残基组成信号肽和后285个氨基酸残基组成成熟肽。
The study was done to obtain a strain which have high lipase production ability and the lipase gene was cloned.Lipase-yield strains were isolated from oily soil samples through plate isolation culture medium with olive oil as substrate while Victoria blue.The lipase activity of isolated strains was assayed by alkaline titrimetric method.The morphologic and physiological-biochemical characteristics of bacteria were preliminarily identified.The 16 S rRNA of bacteria was analyzed.Primers were designed based on the conservative nucleotide sequence and the lipase LipA gene of isolated strain was cloned using genomic DNA as template and analyzed its bioinformatics.The results showed that an isolated bacterial strain number as A05 secreting a large amount of lipase was selected.The lipase activity of strain A05 was assayed 1.07U/ml.The strain A05 was identified as Pseudomonas aeruginosa by testing morphologic and physiological-biochemical characteristics,and the 16 S rRNA sequencing analysis.Primers were designed based on the conservative nucleotide sequence which reported from GenBank,the lipase gene sequence of Pseudomonas aeruginosa A05 was 1092 bp and the lipase gene sequence blast showed 98%identical to the published sequence.Its sequence contains a complete open reading frame,936 bp,encoding 311 amino acids,contained a 26-amino-acid signal sequence and a mature sequence of 285 amino acids.
The study was done to obtain a strain which have high lipase production ability and the lipase gene was cloned.Lipase-yield strains were isolated from oily soil samples through plate isolation culture medium with olive oil as substrate while Victoria blue.The lipase activity of isolated strains was assayed by alkaline titrimetric method.The morphologic and physiological-biochemical characteristics of bacteria were preliminarily identified.The 16 S rRNA of bacteria was analyzed.Primers were designed based on the conservative nucleotide sequence and the lipase LipA gene of isolated strain was cloned using genomic DNA as template and analyzed its bioinformatics.The results showed that an isolated bacterial strain number as A05 secreting a large amount of lipase was selected.The lipase activity of strain A05 was assayed 1.07U/ml.The strain A05 was identified as Pseudomonas aeruginosa by testing morphologic and physiological-biochemical characteristics,and the 16 S rRNA sequencing analysis.Primers were designed based on the conservative nucleotide sequence which reported from GenBank,the lipase gene sequence of Pseudomonas aeruginosa A05 was 1092 bp and the lipase gene sequence blast showed 98%identical to the published sequence.Its sequence contains a complete open reading frame,936 bp,encoding 311 amino acids,contained a 26-amino-acid signal sequence and a mature sequence of 285 amino acids.
引文
[1]张搏,杨江科,闫云君.假单胞菌脂肪酶的研究进展[J].生物技术通报,2007,2:52-57.