Identification of Protease-Producing Halophilic Bacteria Isolated from Salt-Pond Soil (original) (raw)
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Isolation and Identification of Protease Producing Halophilic Bacteria
The protease producing halophilic bacteria were isolated from the banks of different regions of the Indian Ocean near Chennai. In the preliminary screening twelve halophilic bacterial isolates exhibited protease production. Protease enzyme was produced and its activity was assessed through Kunitz assay method. Among them one isolate was taken and identified by Bergey’s manual of systemic bacteriology. The characteristics of the protease producing halophilic Bacillus species HP-2 possess wide range of application in detergent, leather, silver recovery and in food industry. Keywords: : : Protease, Halobacterium, Kunitz assay.
African Journal of Biotechnology, 2017
Gram-positive, rod-shaped, spore-forming haloalkaliphilic bacterium designated as NA7 was isolated from the surface of a Helianthemum nummularium root sample obtained from Wadi Natrun in Egypt. Sequence analysis of the 16S rRNA gene revealed a Bacillus haloalkaliphilius strain as the closest match with 99% identity. In a shake flask culture containing 10% NaCl, adjusted to pH 10 and incubated at 37°C, the isolated strain produced thermostable extracellular alkaline protease with relatively stable maximum activity records (0.610-0.625 TU) within a relatively long stationary phase that exceeded 60 h. A 2-level fractional factorial design (Plackett-Burman) was then applied to screen for nutritional and cultivation factors regulating protease production by the isolate and to appraise their effects. Calculated statistical parameters revealed that NaCl and MgSO 4 are the most significant independent variables affecting alkaline protease production by NA7 and suggested a near-optimum culture condition. Verification of this predicted condition resulted in an alkaline protease specific activity record of 509 TU/mg protein with a 1.27 fold increase when compared to the basal medium culture.
Screening Of A Halotolerant Bacillus Species For Protease Production From Kali Estuary
2024
Uncontrolled anthropogenic activities like urbanization, industrialization, modern agricultural practices and habitat destruction are root cause for generation of pollutants in unprecedented levels. These pollutants are acting mutagens and teratogens causing environmental as well as health hazards. Bacteria as a result of its distribution in various habitats including extremophilic environments evolved their metabolic ability to utilize pollutants and release nonhazardous and useful products in to surroundings by productions of different classes enzymes like laccases, proteases, lipases, hydrolases, dehydrogenases and dehalogenases. In the present study, a mesohalotollerant (6-8%) Bacillus was isolated from Kali estuary by screening on skimmed milk agar media. The identified species was Bacillus vietnamensis. The protease activity was tested against different inducers like peptone, tryptone, milk, groundnut cake and pongamia cake. Among all substrates, peptone showed to be highest inducer of the protease and of the crude protease activity with peptone was showed to be 772.75 U/ml. estimated. The protease is expected to be highly promising for environment clean up, food industry and industrial applications.
SCREENING, ISOLATION AND CHARACTERISATION OF PROTEASE PRODUCING MODERATELY HALOPHILIC MICROORGANISMS
The present study is focused on isolation of moderately halophilic bacteria and fungi that are capable of producing protease from the samples collected from Sambhar lake of Rajasthan and Mumbai seashore. Extremely and moderately halophilic bacteria dominate in saline environments (0.5% to saturated NaCl). Moderately halophilic microorganisms include a broad variety of bacteria and fungi that are able to grow in media containing a wide range of elevated NaCl concentrations (3-15% NaCl). Considerable attention has been focused on enzymes of moderately halophilic bacteria, since they have substantial biotechnological potential. While several proteases from extreme halophiles, members of the haloarchaea have been characterized, fewer proteases from moderately halophilic bacteria have been purified and studied in depth. These microorganisms use different strategies for preserving their cell structure and function in highly saline conditions. They may produce compounds of industrial interest, such as extracellular hydrolytic enzymes with diverse potential applications in the industries. Therefore, an attempt is done in the present study for isolation of moderately halophilic bacteria and fungi able to produce protease which is of industrial importance. The samples collected from Sambhar lake of Rajasthan and Mumbai seashore were processed for the isolation and characterization of the bacteria and fungi able to produce the protease enzyme which further was shown to have the proteolytic activity demonstrated by using blood stained surgical instruments, hair, and casein. The recovery of silver from the X ray photographic film was also attempted. The molecular sequencing of the fungus and bacteria with the phylogenetic tree formulated revealed the presence of Aspergillus flavus in the sample from Mumbai sea shore and Bacillus subtilis from Sambhar lake of Rajasthan. In the subsequent phase effect of temperature, pH, activator and inhibitor was also studied on the activity of protease enzyme by using casein and hair as substrates. The enzyme further was shown to have the ability to remove the blood stain from the surgical instruments as well as to recover silver from the X ray film. The isolate Bacillus subtilis optimally grown at pH-8 and temperature 37°C and 10% NaCl with protease activity on casein and human hair. Protease enzyme obtained in the present study by using Bacillus subtilis and Aspergillus flavus is found to be stable and active so could be of significant use for detergent and leather processing technology as well as for dehairing and silver recovery.
Microorganisms are important source of proteases which are employed in detergent, food, pharmaceutical, leather and photographic industries. Among microorganisms, bacteria have been widely explored globally for their protease production that leads to discovery of several novel proteases. The present research work was carried out to explore the indigenous protease producing soil bacterial flora of Karak, Khyber Pakhtunkhwa, Pakistan. A total of 60 bacteria were isolated from 16 soil samples collected from different sites in district Karak. The isolates were screened for protease production by using skimmed milk agar. Among all the isolates, 18 bacterial isolates were found protease producers with various degree of protease activity. These isolates were further identified through their morphological, microscopic and biochemical examination. It was concluded that soil of Karak is rich in protease producing bacteria that exhibit diversified proteolytic activities and biochemical features. Further optimization of protease activity and their structure elucidation may be performed which will be helpful to identify novel protease.
ISOLATION AND IDENTIFICATION OF PROTEASE PRODUCING BACTERIA FROM SOIL
Protease producing bacteria was isolated from marine soil sample from Marakkanam Saltern, Tamil Nadu. Based on their morphology, it was identified as Pseudonomonas fluorescens biovar. The bacteria were inoculated in gelatin agar for the production of protease enzyme. The crude enzyme extract was purified by ammonium sulphate precipitation method to obtain 85% saturation. The protease enzyme activity of P. fluorescens biovar was 7.5 U/ml after 24 hours of incubation. The maximum extracellular protease production was observed at 37 0 C with a specific activity of 49.02 U/mg.
Water Supply, 2019
An attempt has been made to explore the stability of protease enzyme (isolated from Bacillus sp.) by statistical method. More than 100 isolates were screened for extracellular protease activity derived from various potable water samples of Mahabubnagar district, Telangana State, India. The activity of protease is found to be varying from sample to sample, the highest being reported by the isolate from water sample of Kalwakurthy mandal, Mahabubnagar district and therefore was selected for further studies. The 16S rRNA (ribosomal ribonucleic acid) gene sequence of the isolate showed closest similarity with Bacillus sp. and the sequence was submitted to National Center for Biotechnology Information (NCBI) gene bank (accession number GU566359) and the culture was deposited in three international culture deposition centers (KCTC-13725: MTCC-10465: JCM-16713). The present study revealed that, this Bacillus sp. showed a greater amount of protease production with the inherent characters of...
Isolation and characterization of natural protease producers of Bacillus spp. from Soil samples
2018
Proteases are group of enzymes catalyzing the hydrolysis of proteins. Proteases are considered as commercially most significant among industrial enzymes with wide range of applications in several industries like food, detergent, textile, pulp and paper, and pharmaceutics. Enzymes can be produced by plants, animals or microorganisms, the latter being the dominant host, typically with modified features via recombinant DNA technology for high enzyme production. However, for e.g. food industry, the use of genetically modified microorganisms is not preferred, therefore isolation of new, wild-type microorganism for enzyme production is highly desired. Here we present the results of screening, isolation and characterization of new Bacillus spp. for protease production from soil samples from different areas in Kosovo. Soil samples were divided into four different groups according to their origin: (i) isolates from areas polluted by heavy industry, (ii) isolates from high altitude, (iii) loe...
Screening and Isolation of Extracellular Protease Producing Bacteria from the Maharloo Salt Lake
Iranian Journal of Pharmaceutical Sciences, 2011
Screening and identification of moderately halophilic protease producing bacteria from different regions of Maharloo, a hypersaline lake in the southern area of Iran, were the objectives of this study. In the preliminary screening, 16 isolates exhibited proteolytic activity on saline skim milk agar. All isolates were able to grow comfortably in the media containing 7-15% of salt. Protease activity was assessed through in vitro colorimetric assays for general proteases and the strains were identified by 16S rDNA molecular marker. In comparison to Gram-negative bacteria, the Gram-positive rods, displayed higher proteolytic activities, and Bacillus sp. BCCS 041 was found as the highest protease producing strain with 0.43 U/ml of supernatant activity.
The present study is focused on isolation of moderately halophilic bacteria and fungi that are capable of producing protease from the samples collected from Sambhar lake of Rajasthan and Mumbai seashore. Extremely and moderately halophilic bacteria dominate in saline environments (0.5% to saturated NaCl). Moderately halophilic microorganisms include a broad variety of bacteria and fungi that are able to grow in media containing a wide range of elevated NaCl concentrations (3-15% NaCl). Considerable attention has been focused on enzymes of moderately halophilic bacteria, since they have substantial biotechnological potential. While several proteases from extreme halophiles, members of the haloarchaea have been characterized, fewer proteases from moderately halophilic bacteria have been purified and studied in depth. These microorganisms use different strategies for preserving their cell structure and function in highly saline conditions. They may produce compounds of industrial interest, such as extracellular hydrolytic enzymes with diverse potential applications in the industries. Therefore, an attempt is done in the present study for isolation of moderately halophilic bacteria and fungi able to produce protease which is of industrial importance. The samples collected from Sambhar lake of Rajasthan and Mumbai seashore were processed for the isolation and characterization of the bacteria and fungi able to produce the protease enzyme which further was shown to have the proteolytic activity demonstrated by using blood stained surgical instruments, hair, and casein. The recovery of silver from the X ray photographic film was also attempted. The molecular sequencing of the fungus and bacteria with the phylogenetic tree formulated revealed the presence of Aspergillus flavus in the sample from Mumbai sea shore and Bacillus subtilis from Sambhar lake of Rajasthan. In the subsequent phase effect of temperature, pH, activator and inhibitor was also studied on the activity of protease enzyme by using casein and hair as substrates. The enzyme further was shown to have the ability to remove the blood stain from the surgical instruments as well as to recover silver from the X ray film. The isolate Bacillus subtilis optimally grown at pH-8 and temperature 37°C and 10% NaCl with protease activity on casein and human hair. Protease enzyme obtained in the present study by using Bacillus subtilis and Aspergillus flavus is found to be stable and active so could be of significant use for detergent and leather processing technology as well as for dehairing and silver recovery.