TY  - JOUR
AB  - Corynebacterium glutamicum has been safely used in white biotechnology for the last 60 years and the portfolio of new pathways and products is increasing rapidly. Hence, expression vectors play a central role in discovering endogenous gene functions and in establishing heterologous gene expression. In this work, new expression vectors were designed based on two strategies: (i) a library screening of constitutive native and synthetic promoters and (ii) an increase of the plasmid copy number. Both strategies were combined and resulted in a very strong expression and overproduction of the fluorescence protein GfpUV. As a second test case, the improved vector for constitutive expression was used to overexpress the endogenous xylulokinase gene xylB in a synthetic operon with xylose isomerase gene xylA from Xanthomonas campestris. The xylose isomerase activity in crude extracts was increased by about three-fold as compared to that of the parental vector. In terms of application, the improved vector for constitutive xylA and xylB expression was used for production of the N-methylated amino acid sarcosine from monomethylamine, acetate, and xylose. As a consequence, the volumetric productivity of sarcosine production was 50% higher as compared to that of the strain carrying the parental vector.
DA  - 2021
DO  - 10.3390/microorganisms9010204
KW  - expression vector
KW  - Corynebacterium glutamicum
KW  - overexpression
KW  - promoter
KW  - origin of replication
LA  - eng
IS  - 1
PY  - 2021
T2  - Microorganisms
TI  - Improved plasmid-based inducible and constitutive gene expression in Corynebacterium glutamicum
UR  - https://nbn-resolving.org/urn:nbn:de:0070-pub-29502587
Y2  - 2024-11-22T08:36:24
ER  -