Abstract
Objective: Corynebacterium crenatum AS1.542, a Gram-positive bacterium and indigenous nonpatho-genic corynebacteria, is widely exploited for the in-dustrial production of amino acids. The objective of this paper is to clarify the genetic information of the arginine biosynthetic pathway, and further more contribute to the improvement of arginine produc-tion. Methods: Polymerase chain reaction (PCR) technology was employed for obtaining the arginine biosynthetic gene sequence, and softwares eg. Laser-gene, BPROM, RNAshapes were used for the analysis of obtained sequences. Results: Arginine biosynethetic gene cluster of C. crenatum, comprising argJ, argB, argD, argF, argR and part of argC, has been ampli-fied and sequenced. The gene order has been estab-lished as argCJBDFR, with a entire length of 6.08kb. Conclusion: An internal promoter was found in the upstream of argB gene, four argBDFR ORFs are lo-cated in a same transcription unit, and the tran-scripiton termination of argC gene is irrelevant with the rho-factor. Comparison with ornithine acetyl-transferase (coded by argJ gene) from C. glutamate, ornithine acetyltransferase from C. crenatum also belongs to the monofunctional enzymes.
Highlights
Arginine biosynthesis commences with the acetylation of the amino group of glutamate (Figure 1)
The pathway of arginine biosynthesis can be divided into two parts according to two strategies evolved in the removal of the acetyl group
One is called the “linear” pathway, in which argE gene coded acetylornithinase catalyses the hydrolysis of N-acetylornithine into the arginine precursor ornithine and acetate; the other is called the “economic cyclic” pathway, in which acetylornithine is catalyzed into ornithine and acetyl groups, and recycled with generation of acetylglutamate by argJ gene coded ornithine acetyltransferase [2]
Summary
Arginine biosynthesis commences with the acetylation of the amino group of glutamate (Figure 1). One is called the “linear” pathway, in which argE gene coded acetylornithinase catalyses the hydrolysis of N-acetylornithine into the arginine precursor ornithine and acetate; the other is called the “economic cyclic” pathway, in which acetylornithine is catalyzed into ornithine and acetyl groups, and recycled with generation of acetylglutamate by argJ gene coded ornithine acetyltransferase [2]. The argJ gene by itself would be able to assure both the first and the fifth steps of arginine biosynthesis in above mentioned organisms, there is genetic evidence for the existence of the cloned ornithine acetyltransferase genes from Pseudomonas aeruginosa [4], Saccbaromyces cerevisiae [5], Streptomyces coelicolor [6] and Corynebacterium glutamicum complementing E. coli argE but not argA mutants. The genetic information of arginine biosynthetic pathway was analyzed and clarified in this paper with the aim to contribute to the improvement of arginine production
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