One-step fermentation for producing simvastatin via RNAi silencing of lovF gene in Aspergillus terreus / 生物工程学报

Simvastatin is one of the important prescription drugs for hypercholesterolemia. Monacolin J is a key intermediate during simvastatin synthesis, and also an intermediate of lovastatin biosynthesis. In this work, we construct a monacolin J producing strain via RNA interference to achieve one-step fermentation to obtain simvastatin. The lovF gene silencing plasmid pMHJ137 was constructed and transformed into Aspergillus terreus by Agrobacterium tumefaciens mediated transformation method. Precursor DMB-S-MMP was supplied during the fermentation to screen positive strains of transformants; which also further confirmed the simvastatin producing capability of MJ1-24 by one-step fermentation. Strain MJ1-24 produced monacolin J rather than lovastatin, and the feeding of DMB-S-MMP resulted in the generation of simvastatin. This study suggested that RNAi can efficiently silence the lovF gene of A. terreus and promote the simvastatin production by one-step fermentation.

Biosynthesis of poly (3-mercaptopropionate) and poly (3-mercaptopropionate-co-3-hydroxybutyrate) with recombinant Escherichia coli / 生物工程学报

Polythioesters newly emerged as a type of novel polymer and they have showed great potential for application in industries. In this study, genes of butyrate kinase (buk) and phosphotransbutyrylase (ptb) from Clostridium acetobutylicum, and poly (3-hydroxybutyrate) (PHB) synthase gene from Thiocapsa pfennigii were used for construction of a metabolic pathway to synthesize the polythioesters. When 3-mercaptopropionate and 3-hydroxybutyrate were fed, poly (3-mercaptopropoinate) [poly (3MP)] and poly(3-mercaptopropionate-co-3-hydroxybutyrate) [poly(3MP-co-3HB)] were synthesized by recombinant Escherichia coli JM109 (pBPP1) harboring the constructed metabolic pathway. Results indicated clearly that all these genes are necessary for the synthesis of poly(3MP) and poly(3MP-co-3HB).

Dose-related proconvulsant and anticonvulsant activity of isatin, a putative biological factor, in rats.

Isatin (indole-2, 3-dione) is an endogenous compound with anxiogenic properties, which occur within a narrow dose range (15-20 mg/kg, i.p.). Dose increment beyond 50 mg/kg, i.p. leads to the loss of anxiogenesis. Since a link has been postulated between anxiogenic and convulsant activity, the effect of a range of doses of isatin (20-80 mg/kg, i.p.) was investigated on subconvulsant and convulsant doses of two seizure-inducing agents, namely, pentylenetetrazole (PTZ) and 3-mercapto-propionic acid (3MPA) in rats. Isatin was found to induce a dose-related effect on PTZ and 3MPA convulsions. The lower dose (20 mg/kg, i.p.) potentiated PTZ and 3MPA convulsions, a median dose (40 mg/kg, i.p.) had insignificant effect, whereas higher doses (60 and 80 mg/kg, i.p.) of isatin exhibited significant anticonvulsant effect against both PTZ and 3MPA induced clonic convulsions. The investigation, thus, supports the contention that anxiogenic agents increase the susceptibility to chemical seizures. The proconvulsant effect of isatin, may be due to its inhibitory effect on central atrial natriuretic peptide receptors and stimulation of 5-hydroxytryptamine3 (5-HT3) rather than its monoamine oxidase (MAO) B inhibitory action. The anticonvulsant effect on higher doses of isatin, on the contrary, may be induced by its metabolites, including 5-hydroxyisatin.

Neuropharmacology of amide derivatives of P-GABA.

Three lipophilic amide derivatives of phthaloyl-GABA (P-GABA), namely gamma-phthalimido N-amyl butyramide (PGA), gamma-phthalimido-N-hexylbutyramide (PGH) and gamma-phthalimido N-phenylbutyramide (PGP), were synthesized and evaluated for their hypnotic and anticonvulsant activities in mice. Both PGA and PGH showed moderate hypnotic activity but PGP had no such action. Picrotoxin (0.08 mg/kg) a non-specific GABA antagonist completely abolished the hypnotic action of PGA in subconvulsive doses. Bicuculline (0.04 mg/kg) a GABAA antagonist, 3-mercaptopropionic acid (6 mg/kg) a GAD inhibitor at subconvulsive doses failed to neutralise the hypnotic action of PGA. On the other hand, PGA showed significant protection only against picrotoxin-induced convulsions, but was inactive against other convulsants tested. PGP which has no hypnotic activity, and has a mild anticonvulsant action in all the models except picrotoxin. A definite correlation was observed between the brain GABA and the hypnotic activity of PGA. However the present data indicate that the hypnotic and anticonvulsant activities are mediated probably through different brain GABA-ergic mechanisms.

Antiulcer activity of N-phthaloyl GABA--a new GABA mimetic agent.

N-phthaloyl GABA (P. GABA) inhibited gastric ulceration induced by 3 hr restraint stress at 4 degrees C (CRS) in albino rats. Antiulcer activity of P. GABA was compared with sodium valproate and cimetidine. P. GABA, sodium valproate and cimetidine showed a dose dependent reduction of gastric ulceration. Pretreatment with GABA antagonists-bicuculline methiodide (0.5 mg/kg, im) or 3 mercaptopropionic acid (2 mg/kg, im) reversed the antiulcerogenic activity of both the drugs (P. GABA and sodium valproate). GABA antagonists as such did not induce gastric ulceration in normal rats.