Cloning and expression of a trehalose synthase from Pseudomonas stutzeri CJ38 in Escherichia coli for the production of trehalose.
A novel strain was isolated, Pseudomonas stutzeri CJ38, that enabled direct transformation of maltose to trehalose. In comparison with others reported to date, CJ38 provided a novel trehalose synthase (TSase) without any byproduct, including glucose. Activity analysis, using either maltose or trehalose as a substrate, showed a reversible reaction. There was also no detectable activity of related enzymes with liquid starch and maltooligosaccharides as substrates. Using a malPQ-negative host and MacConkey medium, the TSase gene was cloned in Escherichia coli from CJ38. The resulting sequence contained an open reading frame consisted of 689 amino acids with a calculated molecular mass of 76 kDa. A search for related sequences in various gene and protein data banks revealed a novel family of enzymes that was predicted putatively as a glycosidase or TSase family, with no biochemical evidence. The recombinant enzyme exhibited a high activity toward the substrate maltose, about 50-fold higher than the parent strain and resulted in a high conversion yield (72%) at a relatively high substrate concentration (20%). These results provided the possibility that the strain was effectively used as a potential biocatalyst for the production of trehalose from maltose in a one-step reaction.
Lee
Jin-Ho
JH
R&D Center for Bioproducts, CJ Corporation, Ichon-si, Kyonggi-do 467-812, Korea.
Lee
Kwang-Ho
KH
Kim
Chang-Gyeom
CG
Lee
Se-Young
SY
Kim
Geun-Joong
GJ
Park
Young-Hoon
YH
Chung
Sung-Oh
SO
D004926:Escherichia coli / Q000378:metabolism; D005964:Glucosyltransferases / Q000235:genetics / Q000378:metabolism*; D008320:Maltose / Q000378:metabolism; D008970:Molecular Weight; D015202:Protein Engineering; D044223:Pseudomonas stutzeri / Q000201:enzymology* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D013379:Substrate Specificity; D014199:Trehalose / Q000096:biosynthesis*
D011994:Recombinant Proteins; D008320:Maltose; D014199:Trehalose; D005964:Glucosyltransferases; C099692:trehalose synthase