Trans-4-hydroxy-L-proline(trans-Hyp)is an important chiral compound which can synthesize many pharmaceuticals,and it is also a specific amino acid of collagen.It is widely used in medicine,biochemistry,food and beauty.Although trans-4-hydroxy-L-proline can be synthesized by some microbes directly,yields are very low.At present,hydroxyproline is mainly obtained by acid hydrolysis of collagen,but the raw material is expensive,the separation process is complicated,production efficiency is low and there exist environmental problems.Thus,finding an effective method to produce hydroxyproline is important.Proline-4-hydroxylase is the key enzyme in synthesis of traps-4-hydroxy-L-proline.It can catalyze free L-proline in the presence of 2-oxoglutarate and ferrous ions.In this topic,the genes of tranps-proline-4-hydroxylase(trans-P4H)from diverse resources were cloned and expressed in Corynebacterium glutamicum and Escherichia coli,respectively.The trans-Hyp production by these recombinant strains was investigated.The results showed that all the genes from different resources had been expressed actively.Either the recombinant C.glutamicum or E.coli strains could produce trans-Hyp in the absence of proline and 2-oxoglutarate. |