期刊
BIOTECHNOLOGY LETTERS
卷 35, 期 8, 页码 1265-1270出版社
SPRINGER
DOI: 10.1007/s10529-013-1193-7
关键词
Expression; Phosphoglucomutase; Phosphomannomutase; Sphingan; Sphingomonas sanxanigenens
资金
- Tianjin Research Program of Application Foundation and Advanced Technology of China [11JCZDJC16600]
- Tianjin Science Technology support key Project [10ZCKFSH00900]
- Tianjin Natural Science Foundation [09JCZDJC18000]
- National Natural Science Foundation of China [31000056]
Several strains of the genus Sphingomonas produce sphingans, extracellular polysaccharides used as thickeners, emulsifiers and gelling agents. The pgmG gene from Sphingomonas sanxanigenens, which encodes a bifunctional protein with phosphoglucomutase and phosphomannomutase activities, was cloned and sequenced. The predicted amino acid sequence of the PgmG protein possessed 460 amino acids and a calculated molecular mass of 49.8 kDa, and it was 80 % identical to PGM/PMM from S. elodea. We overexpressed pgmG in Escherichia coli, and the purified protein displayed a K (m) of 0.2 mM and a V (max) of 1.3 mu mol min(-1) mg(-1) with glucose 1-phosphate as substrate. The catalytic efficiency (K (cat)/K (m)) of PgmG was about 15-fold higher for glucose 1-phosphate than for mannose 1-phosphate. Overexpression of pgmG in S. sanxanigenens resulted in a 17 +/- A 0.3 % increase in sphingan production to similar to 12.5 g l(-1).
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