Microbial alkaline proteases: From a bioindustrial viewpoint

The invention includes methods and compositions for remodelling a peptide molecule, including the addition or deletion of one or more glycosyl groups to a peptide, and/or the addition of a modifying group of peptide.

Remodeling and glycoconjugation of peptides

There are basically two kinds of hetero atoms in organic molecules: one kind confers electrophilic character upon the carbon atom to which it is bound, and the other kind turns it into a nucleophilic site. The development of methods permitting transitions between the two resulting categories of reagents has become an important task of modern organic synthesis. The scope of such umpolung of the reactivity of functional groups is discussed for the case of amines as an example. A method of preparing masked α-secondary amino carbanions consists in nitrosation of the secondary amine, followed by metalation of the resulting nitrosamine α-to the nitrogen, reaction with electrophiles, and subsequent denitrosation. Many examples are given for each of these steps which illustrate the wide scope of the overall synthetic operation (electrophilic substitution at the α-C atom of the secondary amine). Preliminary applications and a method for avoiding the handling of nitrosamines are presented, and the report concludes with a brief account of the significance of nitrosamines in the study of carcinogenesis and mutagenesis.

Umpolung of Amine Reactivity. Nucleophilic α-(Secondary Amino)-alkylation via Metalated Nitrosamines†‡

Overproduction of microbial metabolites is related to developmental phases of microorganisms. Inducers, effectors, inhibitors and various signal molecules play a role in different types of overproduction. Biosynthesis of enzymes catalysing metabolic reactions in microbial cells is controlled by well-known positive and negative mechanisms, e.g. induction, nutritional regulation (carbon or nitrogen source regulation), feedback regulation, etc. The microbial production of primary metabolites contributes significantly to the quality of life. Fermentative production of these compounds is still an important goal of modern biotechnology. Through fermentation, microorganisms growing on inexpensive carbon and nitrogen sources produce valuable products such as amino acids, nucleotides, organic acids and vitamins which can be added to food to enhance its flavour, or increase its nutritive values. The contribution of microorganisms goes well beyond the food and health industries with the renewed interest in solvent fermentations. Microorganisms have the potential to provide many petroleum-derived products as well as the ethanol necessary for liquid fuel. Additional applications of primary metabolites lie in their impact as precursors of many pharmaceutical compounds. The roles of primary metabolites and the microbes which produce them will certainly increase in importance as time goes on. In the early years of fermentation processes, development of producing strains initially depended on classical strain breeding involving repeated random mutations, each followed by screening or selection. More recently, methods of molecular genetics have been used for the overproduction of primary metabolic products. The development of modern tools of molecular biology enabled more rational approaches for strain improvement. Techniques of transcriptome, proteome and metabolome analysis, as well as metabolic flux analysis. have recently been introduced in order to identify new and important target genes and to quantify metabolic activities necessary for further strain improvement.

https://sfamjournals.onlinelibrary.wiley.com/doi/pdf/10.1111/j.1751-7915.2007.00015.x

Metabolic regulation and overproduction of primary metabolites

Molecules that interfere with the binding of a tumor necrosis factor receptor with its ligand, such as a soluble receptor, have proven usefulness in both basic research and as therapeutics. The present invention provides improved soluble transmembrane activator and calcium modulator and cyclophilin ligand-interactor (TACI) receptors.

Taci-immunoglobulin fusion proteins

A promoter of a glyceraldehyde-3-phosphate dehydrogenase gene of Acremonium chrysogenum and a transformant which is obtained by using the expression plasmid containing the promoter and is capable of producing a cephalosporin are disclosed

Gylceraldehyde-3-phosphate dehydrogenase gene and promoter

To establish a novel process for the production of L-glutamic acid from n-paraffins, a glycerol auxotroph GL-21, a new type mutant, was successfully obtained from Corynebacterium alkanolyticum No. 314 by treatment with N-methyl-N'-nitro-N-nitrosoguanidine. This auxotroph required glycerol for its growth regardless of the carbon source used. At 72 hr, this mutant GL-21 produced about 40mg/ml of L-glutamic acid from n-paraffins in the culture broth at 0.01 per cent addition of glycerol in the absence of penicillin. A thiamine auxotroph, a biotin auxotroph and an oleic acid auxotroph were also obtained by a similar technique, but these auxotrophs were found to be inapplicable for theproduction of L-glutamic acid from n-paraffins.

https://www.jstage.jst.go.jp/article/bbb1961/36/3/36_3_490/_pdf

Microbial Production of L-Glutamic Acid by Glycerol Auxotrophs Part I

Production of Alkaline Protease from n-Paraffins by a Kabicidin Resistant Mutant Strain of Fusarium sp.

To establish a novel process for the economical production of alkaline protease from n-paraffins by fungi, the variety of fungi were screened for the production of alkaline protease from n-paraffins and Fusarium sp. S–19–5 was selected as an excellent strain, and then the attempt was made to obtain some mutants capable of producing alkaline protease in high yield. A kabicidin resistant mutant No5-128B, was successfully obtained from Fusarium sp. S–19–5 by treatment with ultraviolet light. This mutant No5–128B showed high productivity of alkaline protease, about ten times that of the parent strain. The morphological characteristics and growth response to Kabicidin of the mutant No5–128B were compared with those of the parent stain.

Screening of Fungi Producing Alkaline Protease from n-Paraffins and Induction of Kabicidin Resistant Mutant of Fusarium sp.

To clarify the biosynthetic pathway of maridomycin, the biosynthetic relation among 16-membered macrolide antibiotics belonging to leucomycin-maridomycin group was examined with intact cells or growing cultures of Streptomyces hygroscopicus No. B–5050–HA and its mutants.Carbomycin A, B and leucomycin A3 were converted to maridomycin II in the pH range of 5.5-7.5 with an optimum between 6.0-6.5. But, maridomycin II was not converted to leucomycin A3 or carbomycin B. These findings were confirmed by culture experiment. Leucomycin A3, carbomycin A and B added to the culture at 48 hr were almost completely converted to maridomycin II.On the basis of these facts, we propose a biosynthetic relation among leucomycin A3, carbomycin A, carbomycin B and maridomycin II.

Interconversion among 16-Membered Macrolide Antibiotics Belonging to Leucomycin-Maridomycin Group

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