Microbial production of palatinose through extracellular expression of a sucrose isomerase from Enterobacter sp. FMB-1 in Lactococcus lactis MG1363.
Sucrose isomerase (SIase) has been used to produce palatinose, a structural isomer of sucrose, which has many beneficial health properties, such as low-glycemic and low-insulinemic indices. A gene corresponding to SIase from Enterobacter sp. FMB-1 was expressed in Lactococcus lactis MG1363 using the P170 expression system. The autoinducible promoter (P170) and an optimized signal peptide (SP310mut2) were used to induce and secrete SIase in L. lactis. One-step Ni-NTA affinity chromatography and Western blot analysis demonstrated that SIase was successfully secreted to the culture supernatant, although 60% of the recombinant enzymes were retained inside the cells. The production of the recombinant SIase was highly correlated with pH (pH 6) and glucose concentration (30g/L) of the medium. The extracellularly produced recombinant SIase was functionally active, effectively transforming 50g/L sucrose to 36g/L palatinose, with a conversion rate of 72% in the culture supernatant.
Park
Jong-Yul
JY
Department of Food Science and Biotechnology, Graduate School of Biotechnology and Institute of Life Science and Resources, Kyung Hee University, Yongin, Republic of Korea.
Jung
Jong-Hyun
JH
Seo
Dong-Ho
DH
Ha
Suk-Jin
SJ
Yoon
Jong-Won
JW
Kim
Young-Cheul
YC
Shim
Jae-Hoon
JH
Park
Cheon-Seok
CS
D004754:Enterobacter / Q000502:physiology*; D045604:Extracellular Fluid / Q000201:enzymology; D024861:Genetic Enhancement / Q000379:methods*; D005964:Glucosyltransferases / Q000502:physiology*; D007534:Isomaltose / Q000031:analogs & derivatives* / Q000096:biosynthesis; D013294:Lactococcus lactis / Q000502:physiology*; D011994:Recombinant Proteins / Q000378:metabolism
D011994:Recombinant Proteins; D007534:Isomaltose; D005964:Glucosyltransferases; C116853:sucrose isomerase; C008189:isomaltulose