Kasana Ramesh Chand, Kaur Baljinder, Yadav Sudesh Kumar
Hill Area Tea Science Division, Microbiology Laboratory, Institute of Himalayan Bioresource Technology (CSIR), Palampur, India.
J Basic Microbiol. 2008 Jun;48(3):207-12. doi: 10.1002/jobm.200700160.
Forty three psychrotrophic bacteria were isolated from soil samples collected from Chandra river in sub-alpine region of western Himalaya, India. Among these, 11 isolates were found positive for lipase production at low temperature. Of 11 isolates, CR9 produced largest zone of clearance on plate assay and was able to produce lipase under wide range of pH. The isolate CR9 was identified as Acinetobacter sp. based on morphological and physiochemical characterization and 16S rRNA gene sequencing analysis. According to 16S rRNA gene sequencing data the closest phylogenetic neighbor for strain CR9 was Acinetobacter lwoffii (98.9%). The partially purified lipase from strain CR9 exhibited maximum activity at temperature 40 degrees C and pH optima at 8.0. Cu(2+), Mo(2+), Mg(2+), Zn(2+), phenylmethanesulfonyl fluoride (PMSF), dithiothreitol (DTT) and beta-mercaptoethanol (2-ME) enhanced the enzyme activity, whereas Ca(2+) and ethylenediaminetetraacetic acid (EDTA) had inhibitory effect. Lipase hydrolyzed wide range of short chain fatty acid esters of p-nitrophenyl. The organism CR9 also hydrolyzed tributyrin, Tween 80, soybean oil, mustard oil and olive oil. The results highlight the relevance of unexplored microbes from cold environments of western Himalaya for the isolation of novel lipase producing bacteria.
从印度西喜马拉雅亚高山地区钱德拉河采集的土壤样本中分离出43株嗜冷菌。其中，11株在低温下产脂肪酶呈阳性。在这11株分离菌中，CR9在平板检测中产生的透明圈最大，并且能够在较宽的pH范围内产生脂肪酶。根据形态学、理化特性及16S rRNA基因测序分析，分离菌CR9被鉴定为不动杆菌属。根据16S rRNA基因测序数据，菌株CR9最相近的系统发育邻株是鲁氏不动杆菌（相似度为98.9%）。菌株CR9部分纯化的脂肪酶在40℃时活性最高，最适pH为8.0。铜离子（Cu(2+)）、钼离子（Mo(2+)）、镁离子（Mg(2+)）、锌离子（Zn(2+)）、苯甲基磺酰氟（PMSF）、二硫苏糖醇（DTT）和β-巯基乙醇（2-ME）可增强该酶的活性，而钙离子（Ca(2+)）和乙二胺四乙酸（EDTA）具有抑制作用。脂肪酶可水解多种对硝基苯短链脂肪酸酯。菌株CR9还能水解三丁酸甘油酯、吐温80、大豆油、芥子油和橄榄油。这些结果突出了来自西喜马拉雅寒冷环境中未被探索的微生物对于分离新型产脂肪酶细菌的重要性。
