Cloning and Expression of Rhizopus oryzae of ldhL Gene in Escherichia coli

Abstract:

The ldhL-encoding gene was amplified from Rhizopus oryzae genome DNA using PCR technique, and the PCR product was approximately 1000 bp DNA segment. The PCR products were cloned into pMD18-T vector and identified by enzyme cutting analysis. The sequence results showed that ldhL gene is 963 bp. The DNA fragment of ldhL was subcloned into prokaryotic expression vector pET30a and the specific fusion protein with molecular weight 43 kD was expressed, the result showed that the cloned ldhL was expressed in BL21. The levels of ldhL activity expressed by E. coil BL21/ pET30a-ldhL were up to 98 U/mL. The results are expected to lay foundation for further studies on the gene engineering Rhizopus oryzae strain of high yield L(+)-lactic acid by fermentation of whey.