Construction of lactose-utilizing Xanthomonas campestris and production of xanthan gum from whey.
Xanthomonas campestris pv. campestris possesses a low level of beta-galactosidase and therefore is not able to grow and produce significant amounts of xanthan gum in a medium containing lactose as the sole carbon source. In this study, a beta-galactosidase expression plasmid was constructed by ligating an X. campestris phage phi LO promoter with pKM005, a ColE1 replicon containing Escherichia coli lacZY genes and the lpp ribosome-binding site. It was then inserted into an IncP1 broad-host-range plasmid, pLT, and subsequently transferred by conjugation to X. campestris 17, where it was stably maintained. The lacZ gene under the control of the phage promoter was expressed at a high level, enabling the cells to grow in a medium containing lactose. Production of xanthan gum in lactose or diluted whey by the engineered strain was evaluated, and it was found to produce as much xanthan gum in these substrates as the cells did in a medium containing glucose.
Fu
J F
JF
Department of Botany, National Chung Hsing University, Taichung, Taiwan, Republic of China.
Tseng
Y H
YH
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005503:Food Additives / Q000378:metabolism; D007763:Lac Operon; D007785:Lactose / Q000378:metabolism*; D008894:Milk Proteins / Q000378:metabolism; D010957:Plasmids; D011135:Polysaccharides, Bacterial / Q000378:metabolism; D000067816:Whey Proteins; D014974:Xanthomonas / Q000235:genetics / Q000378:metabolism*; D001616:beta-Galactosidase / Q000235:genetics
D005503:Food Additives; D008894:Milk Proteins; D011135:Polysaccharides, Bacterial; D000067816:Whey Proteins; D001616:beta-Galactosidase; D007785:Lactose; C002563:xanthan gum