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Journal ArticleDOI

Optimization of amylase production using response surface methodology from newly isolated thermophilic bacteria

Wargijono Utomo1
01 Jan 2023-Heliyon (Heliyon)-Vol. 9, Iss: 1, pp e12901-e12901
TL;DR: In this article , the authors aimed at screening and characterizing thermophilic amylase-producing bacteria from water and sediment samples of unexplored hot spring of Tatta Pani Kotli Azad Kashmir.
About: This article is published in Heliyon.The article was published on 2023-01-01 and is currently open access. It has received 3 citations till now. The article focuses on the topics: Thermophile & Amylase.
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Posted ContentDOI
05 May 2023
TL;DR: In this paper , four thermophilic bacteria were screened for thermostable cellulase production on the basis of clear zone formation with 1 % CMC and all the isolates gave maximum activity in presence of CMC upto range of 4.1-4.9 U/mL.
Abstract: Abstract Organic biomass is rich in cellulose and it is the most abundant substrate for biofuel production. To address the challenges posed by cellulosic biomass, cellulolytic microorganisms help improve the feasibility of biofuel production from cellulosic biomass. Present research was aimed to assess the cellulase production and activity of thermophilic isolates discovered from Tata Pani hot spring, Kotli AJ&K. Four thermophilic bacteria were screened for thermostable cellulase production on the basis of clear zone formation with 1 % CMC. Selected strains were Anoxybacillus mongoliensis (MBT001), Anoxybacillus flavithermus (MBT002), Bacillus spp . (MB004) and MBT003 ( Anoxybacillus mongoliensis and Anoxybacillus flavithermus ). Enzyme production and activity was assayed by DNS method. Optimum catalyst formation was observed and validated by RSM at 70 °C, 7 pH and 1.25 % substrate concentration with inocula size of 300 µL after 48 hrs of incubation. All the isolates gave maximum activity in presence of CMC upto range of 4.1-4.9 U/mL. Optimum enzyme activity of crude and purified enzyme was obtained upto range of 3-4 U/mL at pH 7. Crude and purified enzyme was stable upto 70 °C of temperature. Stability of cellulase from selected isolates was varied with surfactants and commercially available detergents being more stable with tween 20 and with commercially available Surf Excel. Cellulase from all isolates exhibited enzymatic hydrolysis of pre-treated banana peels. Crude cellulase from some isolates showed antimicrobial activity against Enterobacter and Pseudomonas . This study has showed that thermophilic bacteria are potential source of thermostable cellulase which can be highly used for many biotechnological purposes.
Journal ArticleDOI
TL;DR: In this article , the authors described the metagenomics-based isolation of a thermostable amylase from the geothermal spring of Odisha using sequence and function-based metagenome.
Abstract: The geothermal springs are said to contain the greatest diversity of undiscovered microorganisms, making them the best source for enzymes with economic significance. The untapped microbial diversity living in the geothermal springs can be mined for novel genes, bioactive substances, and industrially significant biocatalysts using the metagenomics technique. Metagenome was extracted from soil samples of various geothermal springs of India. Metagenome was screened for various carbohydrate degrading enzymes (amylase, cellulase, xylanase, amylopullulanase) using degenerate primers-based Polymerase chain reaction amplifications. Further amplicons were cloned, sequenced and analysis of data was done using various bioinformatics tools, e.g., Blast analysis, Protparam and phre2 program. We have isolated numerous enzymes, including cellulase, amylase, amylopullulanase, and xylanase, from diverse geothermal spring in different parts of India using sequence and function-based metagenomics. In this study, we describe the metagenomics-based isolation of a thermostable amylase from the geothermal spring of Odisha. The amylase gene (1503 bp) was amplified using the metagenome as a template using degenerate primers and cloned into the linearized T vector. The putative gene was likely to encode a protein of 469 amino acids with a molecular weight of 53895.05 Da with pI-7.78. Sequence analysis showed its maximum identity of 98.95% with Bacillus licheniformis alpha-amylase gene. Homology modeling of the amylase protein was done using the phyre2 program, which shows it belongs to the (trans) glycosidase superfamily and contains the catalytic TIM alpha/beta-barrel fold. Hence, we can conclude that geothermal springs are hotspots for the mining of industrially robust biocatalysts.
References
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Journal ArticleDOI
TL;DR: This assay is very reproducible and rapid with the dye binding process virtually complete in approximately 2 min with good color stability for 1 hr with little or no interference from cations such as sodium or potassium nor from carbohydrates such as sucrose.

225,085 citations

Journal ArticleDOI
TL;DR: The latest version of the Molecular Evolutionary Genetics Analysis (Mega) software, which contains many sophisticated methods and tools for phylogenomics and phylomedicine, has been optimized for use on 64-bit computing systems for analyzing larger datasets.
Abstract: We present the latest version of the Molecular Evolutionary Genetics Analysis (Mega) software, which contains many sophisticated methods and tools for phylogenomics and phylomedicine. In this major upgrade, Mega has been optimized for use on 64-bit computing systems for analyzing larger datasets. Researchers can now explore and analyze tens of thousands of sequences in Mega The new version also provides an advanced wizard for building timetrees and includes a new functionality to automatically predict gene duplication events in gene family trees. The 64-bit Mega is made available in two interfaces: graphical and command line. The graphical user interface (GUI) is a native Microsoft Windows application that can also be used on Mac OS X. The command line Mega is available as native applications for Windows, Linux, and Mac OS X. They are intended for use in high-throughput and scripted analysis. Both versions are available from www.megasoftware.net free of charge.

33,048 citations

Journal ArticleDOI
TL;DR: The present review discusses the recent advancement in enzyme technology for food industries, a comprehensive list of enzymes used in food processing, the microbial source of these enzymes and the wide range of their application are discussed.
Abstract: The use of enzymes or microorganisms in food preparations is an age-old process. With the advancement of technology, novel enzymes with wide range of applications and specificity have been developed and new application areas are still being explored. Microorganisms such as bacteria, yeast and fungi and their enzymes are widely used in several food preparations for improving the taste and texture and they offer huge economic benefits to industries. Microbial enzymes are the preferred source to plants or animals due to several advantages such as easy, cost-effective and consistent production. The present review discusses the recent advancement in enzyme technology for food industries. A comprehensive list of enzymes used in food processing, the microbial source of these enzymes and the wide range of their application are discussed.

394 citations

Journal ArticleDOI
TL;DR: An overview of extremophile ecology is provided, and the most recent applications of microbial extremophiles and the extremozymes and extremolytes they produce to biotechnology are reviewed.
Abstract: In the last decade, attention to extreme environments has increased because of interests to isolate previously unknown extremophilic microorganisms in pure culture and to profile their metabolites. Microorganisms that live in extreme environments produce extremozymes and extremolytes that have the potential to be valuable resources for the development of a bio-based economy through their application to white, red, and grey biotechnologies. Here, we provide an overview of extremophile ecology, and we review the most recent applications of microbial extremophiles and the extremozymes and extremolytes they produce to biotechnology.

194 citations

Journal ArticleDOI
TL;DR: A new strictly anaerobic, alkaliphilic, moderately thermophilic, fermentative, spore-forming bacterium, strain K1T, was isolated from manure samples (pH 6-8) and was assigned to the new genus Anoxybacillus gen. nov.
Abstract: A new strictly anaerobic, alkaliphilic, moderately thermophilic, fermentative, spore-forming bacterium, strain K1T, was isolated from manure samples (pH 6-8). Cells were Gram-positive, straight, non-motile rods that grew at temperatures of 37-66 degrees C (optimum at 62 degrees C) and in a pH range of 8.0-10.5 (optimum at 9.5-9.7). The bacterium fermented D-glucose, sucrose, D-fructose, D-trehalose and starch as carbon and energy sources. It required vitamins and its growth is stimulated by yeast extract. The major metabolic products were H2 and acetate. Cells were catalase-negative and could reduce nitrate to nitrite. The G+C content of the DNA was 42.2 mol%. Based on the phenotypic properties and 16S rDNA sequencing and DNA-DNA hybridization data, strain K1T (= DSM 12423T = ATCC 700785T = VKM B-2193T) was assigned to the new genus Anoxybacillus gen. nov., as a representative of a new species, Anoxybacillus pushchinensis sp. nov. 'Bacillus flavothermus' strain d.y., which was found to be closely related to strain K1T, is described as Anoxybacillus flavithermus comb. nov. (type strain = d.y.T = DSM 2641T).

179 citations