Isolation of Eicosapentaenoic Acid-Producing Fungi from Soil Based on Polymerase Chain Reaction Amplification
Isolation of Eicosapentaenoic Acid-Producing Fungi from Soil Based on Polymerase Chain Reaction Amplification
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DOI：
10.5560/znc.2011.66c0429
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发表时间：
2011-07-01
影响因子：
2
通讯作者：
Yu, Long-jiang
中科院分区：
文献类型：
--
作者：
Hu, Jing-rong;Zhou, Peng-peng;Yu, Long-jiang
建立了一种利用聚合酶链反应（PCR）快速筛选和分离产二十碳五烯酸（EPA）土壤真菌的方法。以编码Δ 6脂肪酸去饱和酶和Δ 5脂肪酸去饱和酶的基因为分子标记，从土壤中筛选出产EPA的真菌。65株土壤真菌中有3株PCR扩增阳性。在80 ml马铃薯/葡萄糖液体培养基中于（25 ± 1）℃培养144 h后，发现这三种菌株中有两种产生EPA。EPA产率分别为215.81 mg l（-1）和263.80 mg l（-1）。另一株阳性菌株经检测可产生花生四烯酸（AA）。本研究表明，对Delta 6和Delta 5去饱和酶基因进行分子检测是EPA及其相关多不饱和脂肪酸（PuFAs）产生菌初筛的有效方法，可显著提高初筛效率。
A method was developed for rapid screening and isolation of eicosapentaenoic acid (EPA)-producing soil fungi through polymerase chain reaction (PCR) amplification. Genes coding for Delta 6 fatty acid desaturase and Delta 5 fatty acid desaturase were used as molecular markers for screening these EPA-producing fungi from soil. Three out of 65 soil fungi gave positive results through PCR amplification. Two out of these three strains were found to produce EPA when they had grown in 80 ml potato/dextrose liquid medium at (25 +/- 1) degrees C for 144 h. The EPA yields were 215.81 mg l(-1) and 263.80 mg l(-1), respectively. The other positive strain was detected to produce arachidonic acid (AA). This study indicates that molecular detection of genes encoding Delta 6 and Delta 5 desaturases is an efficient method for primary screening of EPA- or its related polyunsaturated fatty acids (PuFAs)-producing fungi, which can improve the screening efficiency prominently.