| 南极深海沉积物中产低温脂肪酶菌株的筛选与基因克隆 |
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| 引用本文： | 郝文惠,;王凡羽,;郭玉,;汤熙翔. 南极深海沉积物中产低温脂肪酶菌株的筛选与基因克隆[J]. 台湾海峡, 2014, 0(3): 306-311 |
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| 作者姓名： | 郝文惠, 王凡羽, 郭玉, 汤熙翔 |
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| 作者单位： | [1]国家海洋局第三海洋研究所,福建厦门361005; [2]南华大学药学与生命科学院,湖南衡阳421001 |
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| 基金项目： | 南北极环境综合考察与评估专项资助项目（CHINARE2013-01-06）;国家海洋局第三海洋研究所基本科研业务费资助项目（海三科2012003） |
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| 摘 要： | 在10℃恒温培养条件下,从第29次南极科学考察获得的南极深海沉积物样品中分离筛选到产脂肪酶细菌3株,产脂肪酶真菌1株,对其进行分子生物学鉴定,确定为假交替单胞菌属（Pseudoalteromonas arctica）、芽孢杆菌属（Bacillus pumilus）、Glaciecola sp.以及白冬孢酵母属（Leucosporidium sp.）.通过测定各酶的最适反应温度,获得产低温脂肪酶芽孢杆菌XZ18,该菌所产脂肪酶最适反应温度为30℃,在0℃仍有部分活性.从NaCl浓度和pH值2个条件对产酶条件进行研究,最终确定了该菌最适产酶NaCl浓度5.0%、最适产酶pH值为6,最高产酶量为4.65 U/cm3.通过PCR扩增,获得了脂肪酶全长序列,对该序列进行系统发育分析,发现该脂肪酶基因处于较独立的分支.
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| 关 键 词： | 海洋生物学 极地 低温 脂肪酶 基因克隆 |
Screening of cold-active from Antarctic sediments lipase producing strains and cloning of the gene |
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| Affiliation: | HAO Wen-hui, WANG Fan-yu , GUO Yu, TANG Xi-xiang (1. Third Institute of Oceanography, SOA, Xiamen 361005, China; 2. School of Pharmaceutical and Biological Science, University of South China, Hengyang 421001, China) |
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| Abstract: | Three lipase produced bacterial and 1 lipase produced fungi have been screened from Antarctic polar sediments, they are identified as Pseudoalteromonas arctica, Bacillus pumilus, Glaciecola sp. and Leucosporidium sp. by 16S rDNA and ITS DNA sequencing. After detection of the enzyme activity of crude enzyme under different temperatures, we obtain a cold-active lipase producetion strain XZ18 with an optimum temperature of lipase activity at 30% and it remains active at 0℃. By the test of NaCl concentration and pH effects on its lipase production, the optimum conditions of fermentation are found at NaCl concentration 5.0% and pH 6 with the most productive en- zyme quantity at 4.65 U/cm3. With specific designed primers, one lipase gene in XZ18 strain was amplified and sequenced and identified as an independent branch after phylogenetic analysis. |
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| Keywords: | marine biology polar region low-temperature lipase gene clone |
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