• 主题
高级搜索  >
历史搜索 清空历史
首页> 美国卫生研究院文献>Applied and Environmental Microbiology >Thio Wax Ester Biosynthesis Utilizing the Unspecific Bifunctional Wax Ester Synthase/Acyl Coenzyme A:Diacylglycerol Acyltransferase of Acinetobacter sp. Strain ADP1
【2h】

Thio Wax Ester Biosynthesis Utilizing the Unspecific Bifunctional Wax Ester Synthase/Acyl Coenzyme A:Diacylglycerol Acyltransferase of Acinetobacter sp. Strain ADP1

机译：利用非特异性双功能蜡酯合酶/酰基辅酶A：不动杆菌属的二酰基甘油酰基转移酶进行硫蜡酯的生物合成。菌株ADP1

代理获取
本网站仅为用户提供外文OA文献查询和代理获取服务，本网站没有原文。下单后我们将采用程序或人工为您竭诚获取高质量的原文，但由于OA文献来源多样且变更频繁，仍可能出现获取不到、文献不完整或与标题不符等情况，如果获取不到我们将提供退款服务。请知悉。
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

The bifunctional wax ester synthase/acyl coenzyme A (acyl-CoA):diacylglycerol acyltransferase (WS/DGAT) from Acinetobacter sp. strain ADP1 (formerly Acinetobacter calcoaceticus ADP1) mediating the biosyntheses of wax esters and triacylglycerols was used for the in vivo and in vitro biosynthesis of thio wax esters and dithio wax esters. For in vitro biosynthesis, 5′His6WS/DGAT comprising an N-terminal His6 tag was purified from the soluble protein fraction of Escherichia coli Rosetta(DE3)pLysS (pET23a::5′His6atf). By employing SP-Sepharose high-pressure and Ni-nitrilotriacetic acid fast-protein liquid chromatographies, a 19-fold enrichment with a final specific activity of 165.2 nmol mg of protein−1 min−1 was achieved by using 1-hexadecanol and palmitoyl-CoA as substrates. Incubation of purified 5′His6WS/DGAT with 1-hexadecanethiol and palmitoyl-CoA as substrates resulted in the formation of palmitic acid hexadecyl thio ester (10.4% relative specific activity of a 1-hexadecanol control). Utilization of 1,8-octanedithiol and palmitoyl-CoA as substrates led to the formation of 1-S-monopalmitoyloctanedithiol and minor amounts of 1,8-S-dipalmitoyloctanedithiol (59.3% relative specific activity of a 1-hexadecanol control). The latter dithio wax ester was efficiently produced when 1-S-monopalmitoyloctanedithiol and palmitoyl-CoA were used as substrates (13.4% specific activity relative to that of a 1-hexadecanol control). For the in vivo biosynthesis of thio wax esters, the knockout mutant Acinetobacter sp. strain ADP1acr1ΩKm, which is unable to produce fatty alcohols, was used. Cultivation of Acinetobacter sp. strain ADP1acr1ΩKm in the presence of gluconate, 1-hexadecanethiol, and oleic acid in nitrogen-limited mineral salts medium resulted in the accumulation of unusual thio wax esters that accounted for around 1.19% (wt/wt) of the cellular dry weight and consisted mainly of oleic acid hexadecyl thioester as revealed by gas chromatography-mass spectrometry.
机译：来自不动杆菌属的双功能蜡酯合酶/酰基辅酶A（酰基-CoA）：二酰基甘油酰基转移酶（WS / DGAT）。介导蜡酯和三酰基甘油的生物合成的菌株ADP1（以前称为钙乙酸不动杆菌（Acinetobacter calcoaceticus）ADP1）用于硫代蜡酯和二硫代蜡酯的体内和体外生物合成。对于体外生物合成，从大肠杆菌Rosetta（DE3）pLysS（pET23a ::: 5'His6atf）的可溶性蛋白级分中纯化出包含N端His6标签的5'His6WS / DGAT。通过使用SP-琼脂糖高压和Ni-tritrilotriacetic快蛋白液相色谱，富集19倍，最终比活度为165.2 nmol mg蛋白质 -1 min -1 是通过使用1-十六烷醇和棕榈酰辅酶A作为底物实现的。将纯化的5'His6WS / DGAT与1-十六烷硫醇和棕榈酰-CoA作为底物进行孵育导致形成棕榈酸十六烷基硫酯（1-十六烷醇对照的相对比活性为10.4％）。以1,8-辛二硫醇和棕榈酰-CoA为底物导致形成1-S-单棕榈酰泛金丝二硫酚和少量的1,8-S-二棕榈酰泛金丝二硫酚（1-十六烷醇对照的相对比活性为59.3％）。当使用1-S-单棕榈酰泛金刚烷二硫醇和棕榈酰-CoA作为底物时，可以有效地生产后一种二硫蜡酯（相对于1-十六醇对照物的比活性为13.4％）。对于体内生物合成硫蜡酯，敲除突变体不动杆菌属。使用不能产生脂肪醇的菌株ADP1acr1ΩKm。不动杆菌的培养在氮有限的矿物盐培养基中存在葡萄糖酸酯，1-十六烷硫醇和油酸的情况下，菌株ADP1acr1ΩKm导致不寻常的硫代蜡酯积聚，占细胞干重的约1.19％（wt / wt），主要由气相色谱-质谱法显示油酸十六烷基硫酯

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
代理获取

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号