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通过代谢工程改造的谷氨酸棒杆菌生产C50和C40类胡萝卜素及其糖基化

Production and glucosylation of C50 and C 40 carotenoids by metabolically engineered Corynebacterium glutamicum.

作者信息

Heider Sabine A E, Peters-Wendisch Petra, Netzer Roman, Stafnes Marit, Brautaset Trygve, Wendisch Volker F

机构信息

Genetics of Prokaryotes, Faculty of Biology & CeBiTec, Bielefeld University, Universitätsstr. 25, 33615, Bielefeld, Germany.

出版信息

Appl Microbiol Biotechnol. 2014 Feb;98(3):1223-35. doi: 10.1007/s00253-013-5359-y. Epub 2013 Nov 24.

Abstract

The yellow-pigmented soil bacterium Corynebacterium glutamicum ATCC13032 is accumulating the cyclic C50 carotenoid decaprenoxanthin and its glucosides. Carotenoid pathway engineering was previously shown to allow for efficient lycopene production. Here, engineering of C. glutamicum for production of endogenous decaprenoxanthin as well as of the heterologous C50 carotenoids C.p.450 and sarcinaxanthin is described. Plasmid-borne overexpression of genes for lycopene cyclization and hydroxylation from C. glutamicum, Dietzia sp., and Micrococcus luteus, in a lycopene-producing platform strain constructed here, resulted in accumulation of these three C50 carotenoids to concentrations of about 3-4 mg/g CDW. Chromosomal deletion of a putative carotenoid glycosyltransferase gene cg0730/crtX in these strains entailed production of non-glucosylated derivatives of decaprenoxanthin, C.p.450, and sarcinaxanthin, respectively. Upon introduction of glucosyltransferase genes from M. luteus, C. glutamicum, and Pantoea ananatis, these hydroxylated C50 carotenoids were glucosylated. We here also demonstrate production of the C40 carotenoids β-carotene and zeaxanthin in recombinant C. glutamicum strains and co-expression of the P. ananatis crtX gene was used to obtain glucosylated zeaxanthin. Together, our results show that C. glutamicum is a potentially valuable host for production of a wide range of glucosylated C40 and C50 carotenoids.

摘要

黄色色素土壤细菌谷氨酸棒杆菌ATCC13032正在积累环状C50类胡萝卜素脱植基胡萝卜素及其糖苷。此前已证明类胡萝卜素途径工程可实现高效番茄红素生产。本文描述了对谷氨酸棒杆菌进行工程改造以生产内源性脱植基胡萝卜素以及异源C50类胡萝卜素C.p.450和八叠球菌黄素的过程。在此构建的番茄红素生产平台菌株中,通过质粒介导过量表达来自谷氨酸棒杆菌、Dietzia菌属和藤黄微球菌的番茄红素环化和羟基化基因,导致这三种C50类胡萝卜素积累至约3 - 4 mg/g干细胞重量的浓度。在这些菌株中对假定的类胡萝卜素糖基转移酶基因cg0730/crtX进行染色体缺失,分别产生了脱植基胡萝卜素、C.p.450和八叠球菌黄素的非糖基化衍生物。在引入来自藤黄微球菌、谷氨酸棒杆菌和菠萝泛菌的糖基转移酶基因后,这些羟基化的C50类胡萝卜素被糖基化。我们还在此证明了重组谷氨酸棒杆菌菌株中C40类胡萝卜素β-胡萝卜素和玉米黄质的生产,并利用菠萝泛菌crtX基因的共表达获得了糖基化玉米黄质。总之,我们的结果表明谷氨酸棒杆菌是生产多种糖基化C40和C50类胡萝卜素的潜在有价值宿主。

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