Membrane-bound 'synthetic lipase' specifically cultured under solid-state fermentation and submerged fermentation by Rhizopus chinensis: a comparative investigation.
Rhizopus chinensis was able to produce synthetic lipases under both solid-state and submerged fermentations. These lipases were extracted from cell membrane using Triton X-100, and purified to homogeneity through ammonium sulfate precipitation, hydrophobic interaction chromatography and gel filtration chromatography. Judging from SDS-PAGE, the specific synthetic lipases associated with SSF (named as SSL) and SmF (named as SML) were different in the apparent molecular mass (62 and 40kDa). In term of hydrolytic activity, both enzymes exhibited maximum values at pH 8.0 and 40 degrees C; SSL appeared to be more pH tolerant and thermostable than SML. PMSF negligibly affected SSL but strongly reduced the activity of SML. Both enzymes showed clear preference for long-chained p-nitrophenyl esters, yielding maximum activity towards p-nitrophenyl palmitate (with SSL) and p-nitrophenyl laurate (with SML). In term of synthetic activity, lyophilized enzymes gave the highest values both at 30 degrees C, but at different pH memories (7.5 for SSL and 6.5 for SML). Most of ethyl esters synthesized by the two enzymes achieved good yields (>90%), and tetradecanoic acid and laurate acid separately served as the best acyl donors.
Sun
Shu Yang
SY
Key Laboratory of Industrial Biotechnology of Ministry of Education and School of Biotechnology, Jiangnan University, Wuxi, PR China.
Xu
Yan
Y
D019149:Bioreactors / Q000382:microbiology*; D018929:Cell Culture Techniques / Q000379:methods*; D004789:Enzyme Activation; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry; D005285:Fermentation; D008049:Lipase / Q000737:chemistry* / Q000302:isolation & purification; D008567:Membranes, Artificial; D012233:Rhizopus / Q000201:enzymology*; D013379:Substrate Specificity
D004800:Enzymes, Immobilized; D008567:Membranes, Artificial; D008049:Lipase