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Expression of a β‐galactosidase gene from Clostridium acetobutylicum in Lactococcus lactis subsp. lactis
Author(s) -
Pillidge C.J.,
Pearce L.E.
Publication year - 1991
Publication title -
journal of applied bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 0021-8847
DOI - 10.1111/j.1365-2672.1991.tb04666.x
Subject(s) - lactococcus lactis , clostridium acetobutylicum , microbiology and biotechnology , clostridium , gene , chemistry , biology , bacteria , biochemistry , genetics , lactic acid , butanol , ethanol
A β‐galactosidase gene from Clostridium acetobutylicum NCIB 2951 was expressed after cloning into pSA3 and electroporation into derivatives of Lactococcus lactis subsp. lactis strains H1 and 7962. When the clostridial gene was introduced into a plasmid‐free derivative of the starter‐type Lact. lactis subsp. lactis strain H1, the resulting construct had high β‐galactosidase activity but utilized lactose only slightly faster than the recipient. β‐galactosidase activity in the construct decreased by over 50% if the 63 kb Lac plasmid pDI21 was also present with the β‐galactosidase gene. Growth rates of Lac + H1 and 7962 derivatives were not affected after introduction of the clostridial β‐galactosidase, even though β‐galactosidase activity in a 7962 construct was more than double that of the wild‐type strain. When pDI21 was electroporated into a plasmid‐free variant of strain 7962, the recombinant had high phospho‐β‐galactosidase activity and a growth rate equal to that of the H1 wild‐type strain. The H1 plasmid‐free strain grew slowly in T5 complex medium, utilized lactose and contained low phospho‐β‐galactosidase activity. We suggest that β‐galactosidase expression can be regulated by the lactose phosphotransferase system‐tagatose pathway and that Lact. lactis subsp. lactis strain H1 has an inefficient permease for lactose and contains chromosomally‐encoded phospho‐β‐galactosidase genes.