The present investigation highlights the process parameters influencing the submerged fermentation of chitinase by Bacillus pumilus JUBCH08, purification and characterization of the enzyme and determination of antagonistic activity of the bacterium against Fusarium oxysporum. Medium supplemented with 0.5 % chitin and peptone, at initial pH 8.0, when incubated at 35 °C for 72 h favored highest chitinase production. The enzyme was purified 25.1-fold to homogeneity. The chitinase was found to be thermostable and alkali-tolerant with maximum activity at pH 8.0 and 70 °C for 1 h. The molecular weight of chitinase was found to be 64 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Mg2+, Co2+, Ca2+ and Mn2+ improved the chitinase activity. The K m and V max values of the enzyme were 0.13 mg/ml and 38.23 U/ml, respectively. When subjected to dual plate assay, the bacterium showed 45 % antagonism against F. oxysporum. Thus, it could be inferred that cultural conditions strongly affected the chitinase production by B. pumilus JUBCH08. The enzyme being thermostable and best functional under alkaline conditions could be useful for the feed industry and related biotechnological applications. Inhibition of F. oxysporum by the culture through lytic mechanism indicates its potentiality as a biocontrol agent.

/pdf/biosynthesis-and-characterization-of-a-thermostable-alkali-4mvcck1fcv.pdf

Biosynthesis and characterization of a thermostable, alkali-tolerant chitinase from Bacillus pumilus JUBCH08 displaying antagonism against phytopathogenic Fusarium oxysporum

Objective : Aim of this study deals with screening and characterization of chitinase-producing bacteria from marine waste and its deposited soil along the coastal regions in Chennai. Methods : The soil samples were collected aseptically and subjected to serial dilution to isolate the bacterial strains. Totally, 35 morphologically different microorganisms were isolated and were screened for their chitinolytic activity in colloidal chitin incorporated media through zone assay using Congo red stain. The biochemical tests were performed for the isolated to prove their validity and further with sequencing to determine the species. Results: The isolates were screened based on the size of the zone formed. Best chitinase producers were subjected to biochemical tests and 16s ribosomal RNA sequencing. A novel strain, Acinetobacter ASK18, a gram-negative, motile organism was identified. Thus, the isolate may be a potent producer of chitinase, and the marine wastes can be utilized efficiently to generate a high value-added product. Conclusion: A novel strain, Acinetobacter ASK18, would further be subjected to purification of the enzyme produced, and hence the active principle could be evaluated as an effective pharmacological drug in anticancer and antibacterial properties. Keywords : Marine wastes, Chitinase, Congo red, Biochemical tests, 16s rRNA sequencing

Isolation, screening, and characterization of chitinase producing bacteria from marine wastes

In this study, 154 isolates capable of producing extracellular phytate-degrading activity were isolated from four soil samples from volcanic areas in Central Java, Indonesia. Six strains with high phytate-degrading activity were selected for strain identification and characterization of the corresponding phytate-degrading enzyme. Blast analysis of 16S rRNA gene sequences revealed high similarities for all the six isolates to reference sequences belonging to the genusBacillus. Isolates MS5, MC6, D10 and D16 showed 99% sequence identity toB. cereus, while isolate MC8 exhibited 99% sequence identity toB. aryabhatti and D6 99% sequence identity toB. psychrotolerans. The crude extracellular phytase preparations from the isolates showed following optimal conditions for phytate dephosphorylation: pH 4.0 and 50°C (isolate D10), pH 5.0 and 60°C (isolate MC6, and isolate MS5), pH 6.0 and 50°C (isolate D16) and pH 6.0 and 60°C (isolate D6) and pH 6.0 and 40°C (isolate MC8). Zn2+ and Fe3+ strongly inhibited phytate dephosphorylation with all phytase preparations studied. In the presence of Ca2+, an increase in phytase activity of 10-15% was obtained.

/pdf/phytase-producing-bacteria-from-extreme-regions-in-indonesia-vvsp8rw6v8.pdf

Phytase-Producing Bacteria from Extreme Regions in Indonesia

Cold-adapted yeasts include psychrophiles or psychrotolerant nonconventional species able to survive and grow at low temperatures. They represent an important source of biological diversity that has developed a set of structural and functional adaptation strategies to overcome the adverse effects of cold (sometimes associated with other limiting conditions). Among them, the production of cold-active enzymes is probably one of the most efficient adaptations of the eukaryotic physiology at low temperatures. Current literature reports that cold-active enzymes exhibit several advantages than their mesophilic and thermophilic homologues and may successfully replace traditional catalysts in a range of industrial applications carried out at low and moderate temperatures. Due to their singular traits, some cold-active hydrolases (i.e., lipases, amylases, and proteases,) isolated from cold-adapted yeasts have been studied since some decades, while some other, namely, xylanases, chitinases, pectinolytic enzymes, glycosidases, phytases, and invertases, have recently attracted a rising attention for their biotechnological potential from the academy and industry for both food and nonfood exploitations.

Cold-Active Enzymes from Cold-Adapted Yeasts

Crops absorb phosphorous in the form of soluble orthophosphate ion. The solubility of phosphate is inhibited by the presence of iron and aluminium in acidic soils and calcium in neutral and alkaline soils. This leads to fixation of phosphorous, making it not available to crop plants. The phosphate solubilizing bacteria (phosphobacteria) secretes some kinds of organic acids which act on insoluble phosphates and convert the same into soluble form, thus providing phosphorous to plant. An experiment was conducted to enumerate the population density of phosphobacteria in the rhizosphere soils of brinjal, chilly, cotton, green grain, groundnut, maize, paddy, ragi, sorghum and turmeric using Ketznelson and Bose medium following dilution plate technique. Efforts have been made to isolate phosphobacteria from these soils and isolated strains were inoculated in specific media containing specific substrates to produce growth regulating substances such as IAA and GA3 and phosphatase enzyme. The result showed that the population levels of phosphobacteria were higher in the rhizosphere soil of groundnut plant. Further, all the strains of phosphobacteria were able to produce phytohormones and phosphatase enzyme under in vitro conditions.

Short Communication - In vitro production of growth regulators and phosphatase activity by phosphate solubilizing bacteria

Identification and optimization of cultural conditions for chitinase production by Bacillus amyloliquefaciens SM3.

Soil phosphatases play a major role in the mineralization processes of organic phosphorus. A phytase or myo-inositol hexakisphosphate phosphohydrolase (E.C. 3.1.3.8) effectively catalyzes the release of phosphate from phytate and phosphorylated compound. Phytase is derived from different sources such as bacteria and yeasts. The extracellular phytase producing bacteria was isolated from poultry and phytase production was screened. It was purified by ammonium sulphate fractionation of protein and dialysis followed by SDS-PAGE. The DNA from Bacillus megaterium was isolated using CTAB method. The maximum enzyme production was obtained in the 60 th h at 37°C, pH 6.5 and 200 rpm shaking. The best carbon and nitrogen source was lactose 138 µg/mL and ammonium sulphate 124 µg/mL, respectively. The 16S rRNA gene sequencing was carried out for identification of positive bacteria. Pot experiment was carried out with Zea mays for phosphate utilization and accessed by root and maximum measurement was observed.

Screening, Optimization and Application of Extracellular Phytase from Bacillus Megaterium Isolated from Poultry Waste

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Identification and optimization of cultural conditions for chitinase production by Bacillus amyloliquefaciens SM3.

Screening, Optimization and Application of Extracellular Phytase from Bacillus Megaterium Isolated from Poultry Waste