Production of protease by <i>bacillus subtilis</i> using simultaneous control of glucose and ammonium concentrations

Kole, Manoj M.; Draper, I.; Gerson, Donald F.

doi:10.1002/jctb.280410305

Cited by 70 publications

(29 citation statements)

References 13 publications

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“…Interestingly, ammonium chloride, one of the most readily metabolizable nitrogen compound was found to greatly favor the production of protease, being a contrast to the findings reported previously (13,14,22).…”

Section: Isolation and Identification Of The Microorganismcontrasting

confidence: 98%

Isolation, Production, and Characterization of Protease from Bacillus subtilis IB No. 11

Lee¹,

Lee²,

Choi³

et al. 2009

Journal of Animal Science and Technology

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A potent protein degrading bacterium was isolated from soil samples of different environments. Polyphasic taxonomic studies and phylogenetic 16S rRNA sequence analyses led to identify the isolate IB No. 11 as a strain of Bacillus subtilis. The isolated strain was recognized to produce protease constitutively, and the maximum production (1.64 units/ml) was attained in a shake flask culture when the isolate was grown at 40℃, for 32 h in basal medium supplemented with starch (0.25%) and gelatin (1.25%) as sole carbon and nitrogen source, respectively. The optimum pH and temperature for the protease activity were determined to be pH 7.0 and 50℃, respectively. Ca 2+ and Mn 2+ enhanced remarkably the protease activity but neither showed positive effect on the protease's thermal stability. In addition, it was observed that the protease was fairly stable in the pH range of 6.5-8.0 and at temperatures below 50℃, and it could be a good candidate for an animal feed additive. The inhibition profile of the protease by various inhibitors indicated that the enzyme is a member of serine-proteases. A combination of UV irradiation and NTG mutagenesis allowed to develop a protease hyper-producing mutant strain coded as IB No. 11-4. This mutant strain produced approximately 3.23-fold higher protease activity (6.74 units/mg) than the parent strain IB No. 11 when grown at 40℃ for 32h in the production medium. The protease production profile of the selected mutants was also confirmed by the zymography analysis.

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Section: Isolation and Identification Of The Microorganismcontrasting

confidence: 98%

Isolation, Production, and Characterization of Protease from Bacillus subtilis IB No. 11

Lee¹,

Lee²,

Choi³

et al. 2009

Journal of Animal Science and Technology

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“…It is clear from the literature that production of extracellular protease is highly influenced by media components, viz. variation in C/N ratio, presence of easily metabolizable sugars, such as glucose [27][28][29] and presence of metal ions [30]. Besides, several other factors, such as aeration, inoculum density, pH, temperature, and incubation time also affect the protease production [5,31].…”

Section: Resultsmentioning

confidence: 99%

Production and optimization of a commercially viable alkaline protease from a haloalkaliphilic bacterium

Joshi

Dodia

Singh

2008

Biotechnol Bioproc E

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Twenty five haloalkaliphilic bacterial strains were isolated from sea water along the Coastal Gujarat (India) and screened for their ability to secret alkaline proteases. Among them, a potent strain S-20-9 (GenBank accession number EU118360), resembling to Halophilic Bacterium MBIC3303 on the basis of 16S rRNA gene sequencing, was selected for the optimization of enzyme production. S-20-9 produced protease optimally, under aerobic conditions during mid-stationary phase over a broad range of salt (5~25%, w/v) and pH (7~10). The optimum production was at pH 9 and 15% (w/v) NaCl. The production was suppressed by lactose, maltose, sucrose, and inorganic nitrogen sources, especially ammonium ions. Further, the production was significantly stimulated by KH 2 PO 4 and suppressed by glucose. Similarly, the production was also suppressed at higher concentrations of gelatin, yeast extract, peptone, and casamino acids, indicating towards a threshold value for nitrogen requirement. The growth and protease production were enhanced by mono-valent cation (KCl), while the divalent cations acted as inhibitors. The study holds significance as only few reports are available on the alkaline proteases from haloalkaliphilic bacteria, particularly those from moderate saline habitats. © KSBB hÉóïçêÇëW=Ü~äç~äâ~äáéÜáäáÅ=Ä~ÅíÉêá~I=~äâ~äáåÉ=éêçíÉ~ëÉI=éêçÇìÅíáçåI=çéíáãáò~íáçåI=ëÉ~=ï~íÉê= = = = =

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“…The production of extracellular protease in microorganisms is influenced significantly by physical factors such as pH, temperature, dissolved oxygen, incubation time, inoculum level, aeration and agitation and medium constituents (Hanlon et al, 1982;McKeller and Cholette, 1984;Kole et al, 1988;Calik et al, 2002Calik et al, , 2003Shafee et al, 2006;Potumarthi et al, 2007). Response surface methodology (RSM) is a collection of statistical techniques for designing experiments, building models, evaluating the effects of factors and searching optimum levels of parameters for desirable responses (Myers and Montgomery, 2002).…”

Section: Introductionmentioning

confidence: 99%

Effect of physical parameters, carbon and nitrogen sources on the production of alkaline protease from a newly isolatedBacillus pseudofirmus SVB1

2009

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-The effect of process physical parameters (initial pH of the medium, temperature and rpm of the shaking incubator) on the production of alkaline protease from a newly isolated Bacillus pseudofirmus SVB1 was studied using central composite design technique. The individual optimum levels of initial pH of the medium, temperature and rpm of the shaking incubator were found to be 9.2, 27.3 °C and 195, respectively for the production of alkaline protease (specific activity, U/mg of protein) and 11, 28.9 °C and 210, respectively for cell growth. By applying multiresponse analysis method of generalized distance approach for production and growth, the optimal levels were found to be 9.9, 28.1 °C and 202, for initial pH of the medium, temperature and rpm of the shaking incubator, respectively. After optimization, the production of alkaline protease and cell growth were enhanced by 36.23 and 44.29%, respectively. Also, the various nutritional parameters, which have significant effect on the production of alkaline protease, are delineated in the present study. Alkaline protease production was found to be influenced by media components, viz., types of C/N source and presence of metal ions. Finally an overall 6.2 fold increase in the production was achieved using casein as both carbon and nitrogen source.

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Production of protease by bacillus subtilis using simultaneous control of glucose and ammonium concentrations

Cited by 70 publications

References 13 publications

Isolation, Production, and Characterization of Protease from Bacillus subtilis IB No. 11

Isolation, Production, and Characterization of Protease from Bacillus subtilis IB No. 11

Production and optimization of a commercially viable alkaline protease from a haloalkaliphilic bacterium

Effect of physical parameters, carbon and nitrogen sources on the production of alkaline protease from a newly isolatedBacillus pseudofirmus SVB1

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