Biotechnology Division, CSIR - National Institute For Interdisciplinary Science and Technology (NIIST), Trivandrum 695 019, Kerala, India.
Chair of Genetics of Prokaryotes, Bielefeld University, Universitaetsstr. 25, 33615 Bielefeld, Germany.
Bioresour Technol. 2016 Aug;213:239-244. doi: 10.1016/j.biortech.2016.03.019. Epub 2016 Mar 8.
The aim of the present study is the development of a consolidated bioprocess for the production of lysine with recombinant Corynebacterium glutamicum DM1729 strains expressing endoglucanase and β-glucosidase genes. Here, the endoglucanase genes from Xanthomonas campestris XCC3521 and XCC2387 and betaglucosidase gene from Saccharophagus degradans Sde1394 were cloned in C. glutamicum DM1729 and expressed either extracellularly or on cell surface. The highest β-glucosidase activity of 9±0.5U/OD600 of 1 and endoglucanase activity of 5.5±0.8U was obtained in C. glutamicum DM 1729 (pVWEx1-TATXCC2387) (pEKEx3-PorC-Sde1394) when cellobiose (20g/L) alone or in combination with carboxymethyl cellulose (20g/L) was used as the carbon sources respectively. The overall efforts resulted in a lysine titre of 5.9±0.5mM. The ability of the constructs to utilize carboxymethyl cellulose and cellobiose for growth and amino acid production proves the concept of utilization of C. glutamicum as a biocatalyst in the lignocellulosic biorefinery.
本研究的目的是开发一种整合的生物工艺,用于生产赖氨酸,使用表达内切葡聚糖酶和β-葡萄糖苷酶基因的重组棒状杆菌 DM1729 菌株。在这里,从野油菜黄单胞菌 XCC3521 和 XCC2387 中克隆内切葡聚糖酶基因,以及从嗜热脂肪地芽孢杆菌 Sde1394 中克隆β-葡萄糖苷酶基因,并在细胞外或细胞表面表达。当分别使用纤维二糖(20g/L)或与羧甲基纤维素(20g/L)组合作为碳源时,在谷氨酸棒状杆菌 DM1729(pVWEx1-TATXCC2387)(pEKEx3-PorC-Sde1394)中获得了最高的β-葡萄糖苷酶活性 9±0.5U/OD600 和内切葡聚糖酶活性 5.5±0.8U。当单独使用纤维二糖(20g/L)或与羧甲基纤维素(20g/L)组合作为碳源时,在谷氨酸棒状杆菌 DM1729(pVWEx1-TATXCC2387)(pEKEx3-PorC-Sde1394)中获得了最高的β-葡萄糖苷酶活性 9±0.5U/OD600 和内切葡聚糖酶活性 5.5±0.8U。当单独使用纤维二糖(20g/L)或与羧甲基纤维素(20g/L)组合作为碳源时,在谷氨酸棒状杆菌 DM1729(pVWEx1-TATXCC2387)(pEKEx3-PorC-Sde1394)中获得了最高的β-葡萄糖苷酶活性 9±0.5U/OD600 和内切葡聚糖酶活性 5.5±0.8U。总体努力使赖氨酸的产量达到 5.9±0.5mM。这些构建体利用羧甲基纤维素和纤维二糖进行生长和氨基酸生产的能力证明了将谷氨酸棒状杆菌用作木质纤维素生物炼制厂中的生物催化剂的概念。