title: Identification of the genes involved in 1-deoxynojirimycin synthesis in Bacillus subtilis MORI 3K-85.
abstract: 1-Deoxynojirimycin (DNJ), a D-glucose analogue with a nitrogen atom substituting for the ring oxygen, is a strong inhibitor of intestinal α-glucosidase. DNJ has several promising biological activities, including its antidiabetic, antitumor, and antiviral activities. Nevertheless, only limited amounts of DNJ are available because it can only be extracted from some higher plants, including the mulberry tree, or purified from the culture broth of several types of soil bacteria, such as Streptomyces sp. and Bacillus sp. In our previous study, a DNJ-producing bacterium, Bacillus subtilis MORI, was isolated from the traditional Korean fermented food Chungkookjang. In the present study, we report the identification of the DNJ biosynthetic genes in B. subtilis MORI 3K-85 strain, a DNJ-overproducing derivate of the B. subtilis MORI strain generated by γ-irradiation, xhe genomic DNA library of B. subtilis MORI 3K-85 was constructed in Escherichia coli, and clones showing α-glucosidase inhibition activity were selected. After DNA sequencing and a series of subcloning, we were able to identify a putative Operon which consists of gabT1, yktc1, and gutB1 genes predicted to encode putative transaminase, phosphatase, and oxidoreductase, respectively. When a recombinant plasmid containing this Operon sequence was transformed into an E. coli strain, the resulting transformant was able to produce DNJ into the culture medium. Our results indicate that the gabT1, yktc1, and gutB1 genes are involved in the DNJ biosynthetic pathway in B. subtilis MORI, suggesting the possibility of employing these genes to establish a large-scale microbial DNJ overproduction system through genetic engineering and process optimization.
authors: R&D center for Life Science, Biotopia Co., Ltd. Chuncheon, Gangwon-do, 200-881, Republic of Korea.
authors: Kyung-Don
authors: KD
authors: Kang
authors: Yong Seok
authors: YS
authors: Cho
authors: Ji Hye
authors: JH
authors: Song
authors: Young Shik
authors: YS
authors: Park
authors: Jae Yeon
authors: JY
authors: Lee
authors: Kyo Yeol
authors: KY
authors: Hwang
authors: Sang Ki
authors: SK
authors: Rhee
authors: Ji Hyung
authors: JH
authors: Chung
authors: Ohsuk
authors: O
authors: Kwon
authors: Su-Il
authors: SI
authors: Seong
mesh_terms: D017485:1-Deoxynojirimycin / Q000378:metabolism*; D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D065089:Glycoside Hydrolase Inhibitors; D008969:Molecular Sequence Data; D009876:Operon; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D000637:Transaminases / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D065089:Glycoside Hydrolase Inhibitors; D011994:Recombinant Proteins; D017485:1-Deoxynojirimycin; D010088:Oxidoreductases; D000637:Transaminases; D010744:Phosphoric Monoester Hydrolases