Abstract:Proteases are one of the largest groups of hydrolytic enzymes constituting about 60% of total worldwide sales of industrial enzymes due to their wide applications in detergent, leather, textile, food and pharmaceutical industry. Microbial proteases have been preferred over animal and plant proteases because of their fundamental features and ease in production. Bacillus infantis SKS1, an alkaline protease producing bacteria has been isolated from garden soil of north India and identified using morphological, bi… Show more
“…This result, in agreement with other reports ( Joshi et al., 2008 ), indicated that the highest amount of protease was secreted by the both bacterial isolates at alkaline pH. It is also reported that the maximum level of alkaline protease is produced by B. licheniformis MP1 at pH 8.0 ( Jellouli et al., 2011 ) and by B. circulans and B. infantis SKS1 at pH 10.0 ( Prakasham et al., 2005 ; Rao et al., 2009 ; Saggu and Mishra, 2017 ). Fig.…”
Alkaline proteases have applications in numerous industries. In this study, we have isolated and screened proteolytic bacteria from poultry wastes mixed soil and identified two bacterial isolates as Bacillus subtilis AKAL7 and Exiguobacterium indicum AKAL11 based on 16S rDNA sequencing. Maximum level of protease production was achieved after 24 h of fermentation in a basal medium. The optimal temperature, initial pH of the media and agitation for alkaline protease production by these two isolates were 30 °C, pH 9.0 and 120 rpm, respectively. The both bacterial isolates produced maximum level of protease with 3.0% organic municipal solid wastes (OMSW) as the sole source of carbon and nitrogen under previously optimized fermentation conditions. In comparison with the shake flask, protease production increased about 2.5-fold in the bioreactor with reduction in fermentation period. The partial purification of protease resulted in a final 45.67 and 34.86-fold purified protease with a specific activity of 8335.34 and 9918.91 U/mg protein and a typical yield of 9.75 and 9.41% from B. subtilis and E. indicum, respectively. The optimum temperature and pH of the partially purified protease from the both sources was 40 °C and pH 9.0, respectively. Protease from the both isolates was stable at pH 7.0–12.0 and at temperatures up to 50 °C. The effects of protease inhibitors indicated that the protease from B. subtilis might be serine and cysteine type and from E. indicum might be cysteine type. Mg2+, K+ and Ca2+ stimulated but Zn2+, Hg2+, Co2+ and Fe3+ strongly inhibited the protease activity. The partially purified protease from B. subtilis substantially dehaired cow skin and decomposed gelatinous compound from X-ray film. Our study revealed that OMSW can be used as raw material for production of bacterial extracellular protease and alkaline protease from B. subtilis might be potential for industrial and biotechnological applications.
“…This result, in agreement with other reports ( Joshi et al., 2008 ), indicated that the highest amount of protease was secreted by the both bacterial isolates at alkaline pH. It is also reported that the maximum level of alkaline protease is produced by B. licheniformis MP1 at pH 8.0 ( Jellouli et al., 2011 ) and by B. circulans and B. infantis SKS1 at pH 10.0 ( Prakasham et al., 2005 ; Rao et al., 2009 ; Saggu and Mishra, 2017 ). Fig.…”
Alkaline proteases have applications in numerous industries. In this study, we have isolated and screened proteolytic bacteria from poultry wastes mixed soil and identified two bacterial isolates as Bacillus subtilis AKAL7 and Exiguobacterium indicum AKAL11 based on 16S rDNA sequencing. Maximum level of protease production was achieved after 24 h of fermentation in a basal medium. The optimal temperature, initial pH of the media and agitation for alkaline protease production by these two isolates were 30 °C, pH 9.0 and 120 rpm, respectively. The both bacterial isolates produced maximum level of protease with 3.0% organic municipal solid wastes (OMSW) as the sole source of carbon and nitrogen under previously optimized fermentation conditions. In comparison with the shake flask, protease production increased about 2.5-fold in the bioreactor with reduction in fermentation period. The partial purification of protease resulted in a final 45.67 and 34.86-fold purified protease with a specific activity of 8335.34 and 9918.91 U/mg protein and a typical yield of 9.75 and 9.41% from B. subtilis and E. indicum, respectively. The optimum temperature and pH of the partially purified protease from the both sources was 40 °C and pH 9.0, respectively. Protease from the both isolates was stable at pH 7.0–12.0 and at temperatures up to 50 °C. The effects of protease inhibitors indicated that the protease from B. subtilis might be serine and cysteine type and from E. indicum might be cysteine type. Mg2+, K+ and Ca2+ stimulated but Zn2+, Hg2+, Co2+ and Fe3+ strongly inhibited the protease activity. The partially purified protease from B. subtilis substantially dehaired cow skin and decomposed gelatinous compound from X-ray film. Our study revealed that OMSW can be used as raw material for production of bacterial extracellular protease and alkaline protease from B. subtilis might be potential for industrial and biotechnological applications.
“…The enzyme also has a significant role in many physiological and pathological processes i.e. protein catabolism, blood coagulation, zymogen activation and transport and secretion of proteins across membranes [6][7]. The largest application of proteases is in the detergent industry to remove protein-based stains from the cloths.…”
Section: Introductionmentioning
confidence: 99%
“…The enzyme used in the detergent had to be stable in a wide range of temperature and had to be active in the presence of various surfactants, bleaching agents, and various other additives. Currently, a large proportion of commercially available protease is derived from Bacillus strains and most of them were isolated from coastal regions [7][8][9][10][11]. From the study of the previous literature, it can be seen that due to the longest coastal region of Gujarat the possibility of getting the isolate with very good alkaline protease production is high [12][13][14][15][16].…”
Objectives: Microbes from Mangrove ecosystem have the ability to produce useful primary and secondary metabolites of biotechnological significance and hence the objective of this study is to optimize the production of bacterial alkaline protease from a Mangrove isolate Solibacillus silvestris. Methodology: The bacterium (named as Madhwad 10 −3 Summer Zobell) used in the present study was isolated from the soil collected from Madhwad, Gir, India in summer season and was identified using MALDI-TOF (Mass spectrometry). It had shown the highest alkaline protease enzyme activity amongst the other isolates and hence was further analyzed using different physical and chemical factors. Selected parameters i.e. Sucrose, Peptone, and Salinity were further used for the optimization using Response Surface Methodology (RSM). Findings: The bacterium (named as Madhwad 10 −3 Summer Zobell) was identified as Solibacillus silvestris. Among the physical parameters, the results proved that highest enzyme production can be obtained at pH 8.5, 40 °C of temperature, 48 h of incubation period, and agitation speed of 120 rpm while in the chemical parameters, Sucrose and Peptone with 0% salinity were found to give highest enzyme activity i.e. 221.7 U/ml and 246.7 U/ml, respectively. In the statistical analysis done using RSM, all the parameters found to be significant and gave the highest result of 258.750 U/ml at 2.5% Sucrose, 10% Peptone, and 0% Salinity level. Applications/improvements: The enzyme produced due to its efficiency in working at alkaline pH can be utilized in industries such as detergent and textile or in Bioremediation.
“…Proteases are enzymes that catalyze the hydrolysis of peptide bonds, resulting in protein degradation into peptides and amino acids, and also can act in specific posttranslational modification reactions (Souza et al, 2015). Proteolytic enzymes stand out among the enzymes used for industrial applications, representing about 60 % of all commercial enzymes worldwide (Saggu et al, 2017). Proteases can be applied in different industrial sectors, including food industry, detergent additives, textiles, pharmaceuticals, waste treatment, diagnostic reagents, medical application, leather tanning and silver recovery (Marathe et al, 2017;.…”
A strain of Bacillus amyloliquefaciens KUB29 was identified by 16S ribosomal RNA sequencing (Genbank: MF772779.1). Production of thermostable protease, amylase and lipase were done by the isolated strain. The produced enzymes were partially purified by ammonium precipitation followed by dialysis process. Protease and lipase enzymes are effectively used in bio-oil extraction from proteinaceous sample followed by transesterification to produce methyl ester. Amylase enzyme is widely used in food and laundry industry. The produced enzymes are active at thermophilic condition of 55 °C. Use of these enzymes in biofuel production process will make the process cleaner and greener.
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