利用工程大肠杆菌生产不同类型的游离脂肪酸
Production of different types of free fatty acids by engineered E.coli
查看参考文献15篇
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文摘
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游离脂肪酸(free fatty acids,FFAs)对食品风味能够产生重要的影响,生产更多类型的FFAs能够更好的满足食品工业的需求。通过在大肠杆菌(E.coli)中分别过量表达7种不同的硫酯酶基因,7个用于生产FFAs的重组菌株被成功构建。并对重组菌株的FFAs生产条件进行了优化,确定最佳的培养条件为:以15g/L Na_2HPO_4和7.5g/L KH_2PO_4作为培养基中的磷酸盐浓度,采用0.25mmol/L的异丙基硫代-β-D-半乳糖苷(IPTG)在30℃进行诱导培养。在优化的培养条件下,7个重组菌株的FFAs通过气相色谱(GC)进行了定性和定量分析。结果显示,过量表达BTE、AtFatA、‘tesA的重组菌株LL1、LI2、LL4均显著提高了FFAs的产量,其中LL1取得了346.4mg/L最高的FFAs产量。进一步对LL1、LL2、LL4的FFAs组成进行分析表明,BTE、AtFatA、‘TesA具有不同的底物特异性,能够用来生产C12~C18等不同链长和饱和度的FFAs。因此,通过微生物代谢工程可以获得天然动植物油脂中不存在的更多类型的FFAs。 |
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其他语种文摘
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Free fatty acids(FFAs) play a key role in the flavor of many foods.To better meet the needs of flavor industry,it is needed to produce more different types of FFAs.Seven engineered E.coli strains were successfully constructed by overexpressing seven different thioesterases, respectively.Next,taking one engineered strain as an example,an optimization of the production conditions was carried out to improve the production of FFAs. The optimized conditions were 15g/L Na_2HPO_4 and 7.5g/L KH_2PO_4 as the concentration of the phosphates; the engineered strains were induced with 0.25mmol/L IPTG at 30℃.Under this condition,the FFAs production of seven engineered strains was determined by gas chromatography(GC).The strains LL1,LL2 and LL4, which overexpress BTE, AtFatA and 'tesA, respectively, obviously enhanced the production of FFAs, and LL1 achieved 346.4mg/L FFAs, representing the highest FFA titer obtained in all engineered strains.In addition, BTE, AtFatA and 'TesA were found to have different substrate specificities by analyzing the composition of FFAs they achieved, and a range of C12-C18 FFAs was obtained in these engineered strains. Therefore, more types of FFAs, which didn't exist in animal and plant oils,were finally obtained by metabolic engineering of E.coli |
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来源
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食品工业科技
,2012,33(23):158-162 【核心库】
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关键词
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游离脂肪酸
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工程大肠杆菌
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硫酯酶
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地址
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1.
四川农业大学食品科学学院, 四川, 雅安, 625014
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中国科学院青岛生物能源与过程研究所, 山东, 青岛, 266101
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语种
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中文 |
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ISSN
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1002-0306 |
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学科
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轻工业、手工业、生活服务业 |
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基金
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国家自然科学基金
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广东省中国科学院全面战略合作项目
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文献收藏号
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CSCD:4721706
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