Production of Poly(γ-glutamic acid) byBacillus subtilisF-2-01

Purification and Characterization of Poly(γ-glutamic acid) Hydrolase from a Filamentous Fungus,Myrotheciumsp. TM-4222

“…PGA was isolated from 300 liters of the culture supernatant of Bacillus subtilis F-2-01 grown for 3 days at 37°C,21) and was further purified as previously described. 8 ) The purified preparation gave D-and L-glutamic acids at the ratio of 55: 45 after complete acid hydrolysis. Proteinase inhibitors 22 ) such as antipain, chymostatin, and phosphoramidon were products of Meiji Seika Kaisha Co., Ltd. a-L-PGA, poly(a-D-glutamic acid) (a-D-PGA), and l-ethyl-3-(3-dimethylaminopropyl) carbodiimide were purchased from Sigma.…”

Bio-based biodegradable hydrogels prepared by crosslinking of microbial poly(γ-glutamic acid) with L-lysine in aqueous solution

“…3 Modifications of poly(amino acid)s such as PGA have been studied in order to develop high-performance polymers because they are biodegradable and have functional groups. PGA is a polymer produced by some microorganisms such as Bacillus subtilis F-2-01 4 and Bacillus subtilis IFO3335. 5 Crosslinked PGA gels were reported by many researchers for medical or environmental applications, such as drug-release systems [6][7][8][9][10] and water treatment reagents.…”

Genetically Engineered Poly-γ-glutamate Producer fromBacillus subtilisISW1214

“…The MA41 mutant was transformed with these pgs vectors by the competence method. 6) Recombinants were screened on a plate of Luria-Bertani medium 19) with appropriate antibiotics (e.g., Tc, 10 mg ml À1 ; Cm, 5 mg ml À1 ). The recombinant harboring the pWPGS1 vector was tentatively abbreviated to MA41/pWPGS1, and other recombinants constructed were also named in the same way.…”

“…In contrast, the extracellular polymer from Bacillus subtilis is actually a mixture of multi-anionic copolymers in which D-and L-glutamyl residues are randomly aligned, 1) as in the case of poly--DL-glutamate (DL-PGA). Although two distinct mechanisms for the biosynthesis of DL-PGA have been proposed, 1) it was difficult to assess whether, in the polymer production by B. subtilis cells, it is formed from L-glutamate alone, 4,5) as does B. licheniformis, or whether D-glutamate also serves as a substrate 6) according to an amide ligase 7,8) -like manner. 9,10) Intricate regulation of PGA production in B. subtilis 11,12) causes a difficulty in the interpretation of this issue.…”

“…Recent research indicates that the pgsBCA genes encode the sole machinery for PGA synthesis in B. subtilis, 9,16) viz., the PGA synthetase complex, in which the PgsB component shows structural features seen commonly in the amide ligase family 6) but the other two components (PgsC and PgsA) 16) do not encompass y To whom correspondence should be addressed. Fax: +81-888-64-5200; E-mail: ashiuchi@cc.kochi-u.ac.jp Abbreviations: PGA, poly--glutamate; Cm, chloramphenicol; LA-PCR, long amplification-polymerase chain reaction; Tc, tetracycline; SDS-PAGE, SDS-polyacrylamide gel electrophoresis any structural motifs that involve thiotemplate-dependent biopolymer syntheses.…”