Screening and optimization of protease production from a halotolerant Bacillus licheniformis isolated from saltern sediments
TL;DR: Studies on the effect of different carbon and nitrogen sources revealed that xylose and urea enhances the enzyme production, and with selected C–N sources along with 1 M NaCl the maximum protease production was obtained.
About: This article is published in Journal of Genetic Engineering and Biotechnology.The article was published on 2013-06-01 and is currently open access. It has received 67 citations till now. The article focuses on the topics: Bacillus licheniformis & Protease.
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TL;DR: The proteases could be a potential relieves to harmful synthetic chemicals in distinctive industrial processes and thus gains global perception.
99 citations
TL;DR: The protease produced by halo-tolerant Bacillus subtilis strain BLK-1.5 has diverse characteristics and could be useful in various industrial applications.
Abstract: Microbial proteolytic enzyme is one of the most important industrial enzymes that hydrolyze proteins. The applications of proteases under harsh industrial conditions like alkalinity, salinity, and temperature make them inactive and unstable. This suggests need for search for novel microbial sources for protease production having diverse properties. For this purpose, 54 bacterial strains were isolated from different salt mines of Karak, Pakistan and were investigated for their proteolytic activity on skim milk agar plates. The strain which showed maximum protease activity was characterized by 16S rRNA gene sequence analysis. Furthermore, growth and protease production was optimized for the characterized bacteria under different physical factors, i.e., pH, temperature and salinity. The isolate BLK-1.5 exhibited strong protease production and was identified as Bacillus subtilis based on biochemical characteristics and 16S rRNA gene sequence analysis. Maximum production of protease was recorded at pH 10, 37 °C and 7 % (w/v) NaCl. Molecular weight of proteases was estimated 38 kDa and its optimum activity was observed at pH 10, 50 °C and 2 % (w/v) NaCl. In conclusion, the protease produced by halo-tolerant Bacillus subtilis strain BLK-1.5 has diverse characteristics and could be useful in various industrial applications.
43 citations
Cites methods from "Screening and optimization of prote..."
...5 nmol/L phosphate buffer pH 7 and was used for further characterizations (Suganthi et al. 2013)....
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...The precipitated protease was separated by centrifugation at 10,600 rcf for 30 min and the resultant pellet was dissolved in 0.5 nmol/L phosphate buffer pH 7 and was used for further characterizations (Suganthi et al. 2013)....
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TL;DR: The results show that fructose and dextrose serve as the best carbon sources for production of these enzymes, highlighting the use of this strain for enzyme production utilizing relatively inexpensive substrates like beet molasses and corn steep liquor.
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, biochemical and molecular methods. 16S rDNA sequence amplified using universal primers has 99% sequence identity with corresponding gene sequence of Bacillus infantis strain FM 34 and Bacillus sp. Beige. The bacterial culture and its 16S rDNA gene sequence have been deposited to Microbial Culture Collection (Pune, India) with accession number MCC 3035 and GenBank with accession number KR092197 respectively. The partially purified extract of Bacillus infantis SKS1 was thermostable and active in presence of Mg2+, acetyl acetone and laundry detergents implicating its application in industry. Production of these enzymes using this strain was maximized by optimization of various parameters including temperature, pH, media components and other growth conditions. Our results show that fructose and dextrose serve as the best carbon sources for production of these enzymes, highlighting the use of this strain for enzyme production utilizing relatively inexpensive substrates like beet molasses and corn steep liquor. Additionally, this strain showed maximum production of enzymes at 40°C similar to bacterial species used for commercial production of alkaline proteases. Characterization of alkaline proteases from this strain of Bacillus infantis and optimization of parameters for its production would help in understanding its industrial application and large-scale production.
39 citations
TL;DR: Isolation and characterization of each potential actinobacteria having immense industrial and therapeutic value on an unprecedented scale from marine sediments of Visakhapatnam coast will have a burgeoning effect.
32 citations
TL;DR: In this article, a thermostable and wide-range pH stable protease was achieved from a newly isolated Bacillus subtilis K-1 strain using cost-effective agricultural residues.
Abstract: Microbial proteases due to their huge application potential have attracted considerable research attention and account for more than 60% of the worldwide enzyme sales. However, large-scale industrial application of proteases is hindered due to their poor performance under relatively hostile industrial conditions (extremes of temperature, pH) and the high production cost. In the current study, the production of a thermostable and wide-range pH stable protease was accomplished from a newly isolated Bacillus subtilis K-1 strain using cost-effective agricultural residues. Process optimization for protease production was conducted by employing one-variable-at-a-time and statistical approaches. The most significant variables for protease production were identified as incubation time, soybean meal, mustard cake, and wheat bran. Optimization of these variables by central composite design of response surface methodology resulted in a substantial protease yield enhancement (112%). Exploitation of agricultural waste...
30 citations
Cites background from "Screening and optimization of prote..."
...Bacillus strains are known to produce and secrete large quantities of extracellular enzymes and constitute a major group of industrial enzyme producers due to the robust nature of the organism and its enzyme complement (Schallmey et al., 2004; Dijl and Hecker, 2013; Suganthi et al., 2013)....
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...…flour, skimmed milk, and shaker speed 2.6-fold Romsomsa et al. (2010) B. subtilis P13 Soybean meal, incubation time, KH2PO4, FeSO4, and BaCl2 3.2-fold Pillai et al. (2011) B. licheniformis TD 4 Xylose, urea, NaCl, incubation time, pH, and agitation rate 1.57-fold Suganthi et al. (2013) Bacillus sp....
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References
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TL;DR: The moderately halophilic heterotrophic aerobic bacteria form a diverse group of microorganisms and approaches to the study of genetic processes have recently been developed, opening the way toward an understanding of haloadaptation at the molecular level.
Abstract: The moderately halophilic heterotrophic aerobic bacteria form a diverse group of microorganisms. The property of halophilism is widespread within the bacterial domain. Bacterial halophiles are abundant in environments such as salt lakes, saline soils, and salted food products. Most species keep their intracellular ionic concentrations at low levels while synthesizing or accumulating organic solutes to provide osmotic equilibrium of the cytoplasm with the surrounding medium. Complex mechanisms of adjustment of the intracellular environments and the properties of the cytoplasmic membrane enable rapid adaptation to changes in the salt concentration of the environment. Approaches to the study of genetic processes have recently been developed for several moderate halophiles, opening the way toward an understanding of haloadaptation at the molecular level. The new information obtained is also expected to contribute to the development of novel biotechnological uses for these organisms.
1,266 citations
TL;DR: This review focuses on the various options that may be employed to improve microbial strains and addresses the complex problems of screening, the tools and technology behind the selection of targeted organisms, and the importance of process optimization.
Abstract: Improvement of microbial strains for the over-production of industrial products has been the hallmark of all commercial fermentation processes. Conventionally, strain improvement has been achieved through mutation, selection, or genetic recombination. Over-production of primary or secondary metabolites is a complex process, and successful development of improved strains requires a knowledge of physiology, pathway regulation and control, and the design of creative screening procedures. In addition, it requires mastery of the fermentation process for each new strain, as well as sound engineering know-how for media-optimization and the fine-tuning of process conditions. This review focuses on the various options that may be employed to improve microbial strains and addresses the complex problems of screening, the tools and technology behind the selection of targeted organisms, and the importance of process optimization. Furthermore, this review discusses new and emerging technologies and designing optimized media for tracking mutants with enhanced productivity or other desired attributes.
499 citations
TL;DR: Despite the unmistakable organizational similarities between the B. licheniformis and B. subtilis genomes, there are notable differences in the numbers and locations of prophages, transposable elements and a number of extracellular enzymes and secondary metabolic pathway operons that distinguish these species.
Abstract: Bacillus licheniformis is a Gram-positive, spore-forming soil bacterium that is used in the biotechnology industry to manufacture enzymes, antibiotics, biochemicals and consumer products. This species is closely related to the well studied model organism Bacillus subtilis, and produces an assortment of extracellular enzymes that may contribute to nutrient cycling in nature. We determined the complete nucleotide sequence of the B. licheniformis ATCC 14580 genome which comprises a circular chromosome of 4,222,336 base-pairs (bp) containing 4,208 predicted protein-coding genes with an average size of 873 bp, seven rRNA operons, and 72 tRNA genes. The B. licheniformis chromosome contains large regions that are colinear with the genomes of B. subtilis and Bacillus halodurans, and approximately 80% of the predicted B. licheniformis coding sequences have B. subtilis orthologs. Despite the unmistakable organizational similarities between the B. licheniformis and B. subtilis genomes, there are notable differences in the numbers and locations of prophages, transposable elements and a number of extracellular enzymes and secondary metabolic pathway operons that distinguish these species. Differences include a region of more than 80 kilobases (kb) that comprises a cluster of polyketide synthase genes and a second operon of 38 kb encoding plipastatin synthase enzymes that are absent in the B. licheniformis genome. The availability of a completed genome sequence for B. licheniformis should facilitate the design and construction of improved industrial strains and allow for comparative genomics and evolutionary studies within this group of Bacillaceae.
439 citations
TL;DR: The production of alkaline protease was optimized using a newly isolated Bacillus sp.
308 citations
TL;DR: A newly isolated haloalkaliphilic Bacillus sp.
214 citations