Optimization of medium and cultivation conditions forl-amino acid oxidase production byAspergillus fumigatus

2009 ◽  
Vol 55 (9) ◽  
pp. 1096-1102 ◽  
Author(s):  
Susmita Singh ◽  
B. K. Gogoi ◽  
R. L. Bezbaruah

A fungal strain was selected from the microbial repository of the North-East Institute of Science and Technology, Jorhat, India, which could produce a high yield of l-amino acid oxidase. 18SrRNA, ITS1, 5.8SrRNA ITS2, and partial 28 S rRNA sequencing and phenotypic characteristics indicate that it belong to the species Aspergillus fumigatus (designated as P13). Maximum production of enzyme (59.55 × 10−3 U/mg dry cell mass) was obtained in a medium containing 10 g/L glucose, 4 g/L yeast extract, and 4 g/L ammonium sulfate, with 20 mmol/L of l-threonine as the inducer. The optimum temperature for enzyme production was 30 °C at pH 7.0, with a shaking speed of 200 r/min. At 96 h, the enzyme activity was maximum. The A. fumigatus P13 l-amino acid oxidase accepts a broad substrate range, and the maximum enzyme activity (20.41 × 10−3 U/mg dry cell mass) was obtained with 50 mmol/L of dl-tyrosine. In the literature, no reports have been found regarding the production of l-amino acid oxidase by A. fumigatus. The enzyme showed enantiomerically pure amino acid formation, which has tremendous demand in industrial applications.

1997 ◽  
Vol 43 (3) ◽  
pp. 292-295 ◽  
Author(s):  
Salim K. Mujawar ◽  
Jaiprakash G. Shewale

Aspergillus sp. strain O20 produces inducible D-amino acid oxidase intracellularly, only in the presence of some amino acids. The enzyme was induced most effectively by the addition of DL-alanine (1% w/v) to the production medium. Among the various compounds studied, production of the D-amino acid oxidase was enhanced by Aerosol-22, glucose, and sodium nitrate. D-Amino acid oxidase formation was observed during the onset of the stationary growth phase. Maximum enzyme activity was recorded after 96 h of fermentation (1000 IU/L).Key words: D-amino acid oxidase, Aspergillus sp., 7-aminocephalosporanic acid, cephalosporin C.


2013 ◽  
Vol 41 (6) ◽  
pp. 1551-1556 ◽  
Author(s):  
Silvia Sacchi

Over the years, accumulating evidence has indicated that D-serine represents the main endogenous ligand of NMDA (N-methyl-D-aspartate) receptors. In the brain, the concentration of D-serine stored in cells is defined by the activity of two enzymes: serine racemase (responsible for both the synthesis and degradation) and D-amino acid oxidase (which catalyses D-serine degradation). The present review is focused on human D-amino acid oxidase, discussing the mechanisms involved in modulating enzyme activity and stability, with the aim to substantiate the pivotal role of D-amino acid oxidase in brain D-serine metabolism.


1989 ◽  
Vol 261 (1) ◽  
pp. 233-238 ◽  
Author(s):  
K Zaar ◽  
A Völkl ◽  
H D Fahimi

D-Aspartate oxidase (EC 1.4.3.1) was assayed in subcellular fractions and in highly purified peroxisomes from rat, bovine and sheep kidney cortex as well as from rat liver. During all steps of subcellular-fractionation procedures, D-aspartate oxidase co-fractionated with peroxisomal marker enzymes. In highly purified preparations of peroxisomes, the enrichment of D-aspartate oxidase activity over the homogenate is about 32-fold, being comparable with that of the peroxisomal marker enzymes catalase and D-amino acid oxidase. Disruption of the peroxisomes by freezing and thawing released more than 90% of the enzyme activity, which is typical for soluble peroxisomal-matrix proteins. Our findings provide strong evidence that in these tissues D-aspartate oxidase is a peroxisomal-matrix protein and should be added as an additional flavoprotein oxidase to the known set of peroxisomal oxidases.


Genetics ◽  
1983 ◽  
Vol 103 (2) ◽  
pp. 277-285 ◽  
Author(s):  
Ryuichi Konno ◽  
Yosihiro Yasumura

ABSTRACT d-Amino acid oxidase activity in the kidney homogenates of mice of seven strains was measured to search for a mutant for this enzyme. There was a consistent sex difference in the enzyme activity in these strains: male mice showed higher levels of the enzyme activity than females. In contrast to other strains, some mice of the ddY strain did not possess enzyme activity. This trait was inheritable, and a mouse stock without enzyme activity (DAO-) was established. The allele (Dao-1c) carried by the DAO- mice was recessive and behaved as a single autosomal gene in inheritance. Heterozygous mice for this gene (Dao-1  +/Dao-1c) showed nearly half the enzyme activity of the wild-type homozygotes (Dao-1  +/Dao-1  +), suggesting that Dao-1c is a null allele and that there is a gene dosage effect on the enzyme activity.


Author(s):  
Charles D. Derby ◽  
Eric S. Gilbert ◽  
Phang C. Tai

Many marine animals use chemicals to defend themselves and their eggs from predators. Beyond their ecologically relevant functions, these chemicals may also have properties that make them beneficial for humans, including with biomedical and industrial applications. For example, some chemical defenses are also powerful antimicrobial or anti-tumor agents with relevance to human health and disease. One such chemical defense, Escapin, an L–amino acid oxidase in the defensive ink of the sea hare Aplysia californica, and related proteins have been investigated for their biomedical properties. This review details our current understanding of Escapin’s antimicrobial activity, including the array of chemicals generated by Escapin’s oxidation of its major substrates, L–lysine and L–arginine, and mechanisms underlying these molecules’ bactericidal and bacteriostatic effects on planktonic cells and the prevention of formation and removal of bacterial biofilms. Models of Escapin’s effects are presented, and future directions are proposed.


2008 ◽  
Vol 74 (11) ◽  
pp. 3400-3409 ◽  
Author(s):  
Chiaki Yamada ◽  
Kyoko Kijima ◽  
Sayaka Ishihara ◽  
Chinatsu Miwa ◽  
Kei Wada ◽  
...  

ABSTRACT 7-Aminocephalosporanic acid (7-ACA) is an important material in the production of semisynthetic cephalosporins, which are the best-selling antibiotics worldwide. 7-ACA is produced from cephalosporin C via glutaryl-7-ACA (GL-7-ACA) by a bioconversion process using d-amino acid oxidase and cephalosporin acylase (or GL-7-ACA acylase). Previous studies demonstrated that a single amino acid substitution, D433N, provided GL-7-ACA acylase activity for γ-glutamyltranspeptidase (GGT) of Escherichia coli K-12. In this study, based on its three-dimensional structure, residues involved in substrate recognition of E. coli GGT were rationally mutagenized, and effective mutations were then combined. A novel screening method, activity staining followed by a GL-7-ACA acylase assay with whole cells, was developed, and it enabled us to obtain mutant enzymes with enhanced GL-7-ACA acylase activity. The best mutant enzyme for catalytic efficiency, with a k cat/Km value for GL-7-ACA almost 50-fold higher than that of the D433N enzyme, has three amino acid substitutions: D433N, Y444A, and G484A. We also suggest that GGT from Bacillus subtilis 168 can be another source of GL-7-ACA acylase for industrial applications.


1956 ◽  
Vol 185 (2) ◽  
pp. 250-256 ◽  
Author(s):  
Barbara R. Endahl ◽  
Charles D. Kochakian

A large number of C19 steroids were able to markedly increase the d-amino acid oxidase activity of the kidney of the castrated mouse. The maximum effect was attained within 21 days of treatment and with relatively small doses of the most potent androgens. On the other hand, neither castration nor various androgens were able to significantly alter the d-amino acid oxidase activity of either the kidney or liver of the castrated rat and guinea pig. Furthermore, age did not influence the enzyme activity of the tissues of the rat (2–8 months of age) or the guinea pig (4–8 months of age). Estradiol produced a small increase in the d-amino acid oxidase activity of the kidney of the mouse but estrone, methoxybisdehydrodiosynolic acid and several corticoids were ineffective.


The oxidation of D-alanine, hypoxanthine and glucose by D-amino-acid oxidase, xanthine oxidase and glucose oxidase (notatin) respectively has been studied at O 2 tensions varying from 1 to 100 %, mainly with a view to examining enzyme activity at an O 2 tension comparable to that obtaining in the animal body, i.e. < 10 % O 2 . It was found that ( a ) the rate of substrate oxidation decreases markedly at O 2 tensions < 20 % , i.e. the enzymes examined have a low O 2 affinity; ( b ) the substrate concentration giving half-maximal oxidation rate (= Michaelis constant, K m ) decreases with a lowering of the O 2 tension; ( c ) the rate of primary substrate oxidation at low O 2 tension in presence of catalase is higher than in presence of catalase and a secondary substrate which undergoes peroxidatic oxidation by catalase and H 2 O 2 . A possible mechanism underlying this effect, which involves the participation of catalase and H 2 O 2 in primary substrate oxidation at low O 2 tension, is suggested.


2021 ◽  
Author(s):  
Jian-Miao Xu ◽  
Hui-Ting Cao ◽  
Ming Wang ◽  
Bao-Jian Ma ◽  
Liu-Yu Wang ◽  
...  

Abstract d-amino acid oxidase (DAAO) is widely used in the industrial preparation of l-amino acids, and cultivating Escherichia coli (E. coli) expressing DAAO for the biosynthesis of l-phosphinothricin (l-PPT) is very attractive. At present, the biomass production of DAAO by fermentation is still limited in large-scale industrial applications because the expression of DAAO during the fermentation process inhibits the growth of host cells, which limits higher cell density. In this study, the factors that inhibit the growth of bacterial cells during a 5 L fed-batch fermentation process were explored, and the fermentation process was optimized by co-expressing catalase (CAT), by balancing the biomass and the enzyme activity, and by adding exogenous d-alanine (d-Ala) to relieve the limitation of DAAO on the cells and optimize fermentation. Under optimal conditions, the DO-STAT feeding mode with DO controlled at 30% ± 5% and the addition of 27.5 g/L lactose mixed with 2 g/L d-Ala during induction at 28 °C resulted in the production of 26.03 g dry cell weight (DCW)/L biomass and 390.0 U/g DCW specific activity of DAAO; an increase of 78% and 84%, respectively, compared with the initial fermentation conditions. The fermentation strategy was successfully scale-up to a 5000L fermenter.


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