Biochemical properties and repression studies of an alkaline serine protease from a haloalkaliphilic actinomycete, Nocardiopsis dassonvillei subsp. albirubida OK-14

Dwivedi, Purna; Sharma, Amit Kumar; Singh, Satya P.

doi:10.1016/j.bcab.2021.102059

Cited by 8 publications

(4 citation statements)

References 46 publications

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“…The characterization method of the influence of pH value on the activity and stability had referred to the report of Tito et al and Dwivedi et al [ 29 , 30 ], and was appropriately modified in this study.…”

Section: Materials and Methodologymentioning

confidence: 99%

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Development of a microbial protease for composting swine carcasses, optimization of its production and elucidation of its catalytic hydrolysis mechanism

Zhai

Duan

et al. 2022

BMC Biotechnol

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Background Dead swine carcass composting is an excellent method for harmless treatment and resource utilization of swine carcass. However, poor biodegradation ability of traditional composting results in poor harmless treatment effect. Researches report that the biodegradation ability of composting can be improved by inoculation with enzyme-producing microorganisms or by inoculation with enzyme preparations. At present, the researches on improving the efficiency of dead swine carcass composting by inoculating enzyme-producing microorganisms have been reported. However, no work has been reported on the development of enzyme preparations for dead swine carcass composting. Methodology The protease-producing strain was isolated by casein medium, and was identified by 16 S rRNA gene sequencing. The optimal fermentation conditions for maximum protease production were gradually optimized by single factor test. The extracellular protease was purified by ammonium sulfate precipitation and Sephadex G-75 gel exclusion chromatography. The potential for composting applications of the purified protease was evaluated by characterization of its biochemical properties. And based on amino acid sequence analysis, molecular docking and inhibition test, the catalytic hydrolysis mechanism of the purified protease was elucidated. Results In this study, a microbial protease was developed for swine carcass composting. A protease-producing strain DB1 was isolated from swine carcass compositing and identified as Serratia marcescen. Optimum fermentation conditions for maximum protease production were 5 g/L glucose, 5 g/L urea, 1.5 mmol/L Mg2+, initial pH-value 8, inoculation amount 5%, incubation temperature 30 °C and 60 h of fermentation time. The specific activity of purified protease reached 1982.77 U/mg, and molecular weight of the purified protease was 110 kDa. Optimum pH and temperature of the purified protease were 8 and 50 °C, respectively, and it had good stability at high temperature and in alkaline environments. The purified protease was a Ser/Glu/Asp triad serine protease which catalyzed substrate hydrolysis by Glu, Arg, Ser, Asp and Tyr active residues. Conclusions In general, the microbial protease developed in this study was suitable for industrial production and has the potential to enhance composting at thermophilic stage. Moreover, the catalytic hydrolysis mechanism of the protease was further analyzed in this study.

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Section: Materials and Methodologymentioning

confidence: 99%

“…Based on the experiment results of optimum metal ion, incubation temperatures were set to 20,25,30,35,40,45 °C. The activities of crude proteases were determined to determine the optimum incubation temperature.…”

Section: Optimum Incubation Temperaturementioning

confidence: 99%

Development of a microbial protease for composting swine carcasses, optimization of its production and elucidation of its catalytic hydrolysis mechanism

Zhai

Duan

et al. 2022

BMC Biotechnol

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“…Enzymes from haloalkaliphilic microorganisms have gained considerable interest in recent years due to their excellent stability and activity in high salt, pH, and temperature (Bhatt and Singh, 2020). Recent studies on the extracellular enzymes of the microorganisms dwelling in extreme habitats of high salinity and alkaline pH have established their ecological and biotechnological significance (Purohit and Singh, 2011;Sinha and Khare, 2013;Raval et al, 2015;Baweja et al, 2016;Raval et al, 2018;Si et al, 2018;Thakrar and Singh, 2019;Bhatt and Singh, 2020;Dwivedi et al, 2021;Rathore and Singh, 2021).…”

Section: Introductionmentioning

confidence: 99%

Diversity of Cultivable Bacteria in A Saline Desert of Little Rann of Kutch, India: A Phylogenetic Perspective

Bhatt

Singh

2022

Front. Mar. Sci.

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Deserts in general, and Indian deserts in particular, are less attended for microbial diversity. The Little Rann of Kutch (LRK), a coastal saline desert, is characterized by a unique combination of both dry and wet features. This study represents the first report on the extensive isolation, spatial distribution, 16S rRNA gene-based phylogeny, and identification of novel taxa. A total of 87 isolates were obtained from three different study sites in LRK. Based on the full 16S rRNA gene sequences, the isolates were grouped into 44 different phylotypes of four phyla: Firmicutes, Proteobacteria, Actinobacteria, and Euryarchaeota. These in turn were represented by 19 different genera. Halomonas, Gracilibacillus, Thalassobacillus, Piscibacillus, Salimicrobium, Alkalibacillus, Bhargavaea, Proteus, Marinobacter, Pseudomonas, Kocuria, Corynebacterium, Planococcus, Micrococcus and Natronococcus identified in this study had never before been reported from this habitat. A majority of the isolates displayed broad salt and pH tolerance. The bacterial diversity of Venasar and Jogad closely resembled with each other. While Bacillus, Virgibacillus, Gracillibacillus, and Bhargavaea were common genera in all sites, six putative novel taxa of different phylogenetic groups were identified. Available nitrogen, pH, Organic carbon, TDS, and EC were the main environmental variables affecting the microbial diversity. Analysis of the geographical distribution revealed that a majority of the phylotypes had cosmopolitan distribution, followed by the saline and marine distribution, while ∼13% were affiliated with only LRK. The phylotypes associated with marine distribution decreased with increasing distance from the Gulf of Kutch, suggesting their endemism to marine environments. The study established the taxonomic novelty and prospects for the discovery of unique products and metabolites.

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“…Proteolytic enzymes are used both in the processing of by-products of meat, dairy and poultry processing industry, and when developing food products. According to Dwivedi et al [5], purified extracellular keratinase obtained from Aspergillus (Aspergillus sp. DHE7) shows its activity against casein, bovine serum albumin, gelatin, to feathers and wool in the pH range of 7-11 at a temperature of 37-70 • C. The paper of El-Ghonemy and Ali [6] provides data on the study of the alkaline protease Nocardiopsis dassonvillei subsp.…”

Section: Introductionmentioning

confidence: 99%

Efficiency of Microencapsulation of Proteolytic Enzymes

et al. 2021

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Currently, special attention is paid to the study of the effectiveness of the immobilization method—microencapsulation. The aim of the research is to obtain a complex enzyme preparation from pepsin and papain by sequential microencapsulation of enzymes in a pseudo-boiling layer and to evaluate its tenderizing effect on pork. The objects of research were enzymes: pepsin and papain, which were microencapsulated in a protective coating of maltodextrin. It was found that the biocatalytic activity of the complex enzyme preparation is higher than that of pure enzymes. Microencapsulation allows maintaining the high proteolytic activity of enzymes for a long storage period. It has been shown that the thickness of the protective layer during microencapsulation of pepsin and papain in the pseudo-boiling layer of maltodextrin should be in the range of 4–6 microns. During the research, the physicochemical properties of pork were studied depending on the duration of fermentation. It was found that the maximum activity of immobilized enzymes is shifted to the alkaline side. Pork salting with the use of a microencapsulated enzyme preparation in the brine increases the water-binding capacity of proteins to a greater extent in comparison with brine with pure enzymes. The presented data show the high efficiency of sequential microencapsulation of the enzyme pepsin and then papain into a protective layer of maltodextrin in order to preserve their activity during storage.

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Biochemical properties and repression studies of an alkaline serine protease from a haloalkaliphilic actinomycete, Nocardiopsis dassonvillei subsp. albirubida OK-14

Cited by 8 publications

References 46 publications

Development of a microbial protease for composting swine carcasses, optimization of its production and elucidation of its catalytic hydrolysis mechanism

Development of a microbial protease for composting swine carcasses, optimization of its production and elucidation of its catalytic hydrolysis mechanism

Diversity of Cultivable Bacteria in A Saline Desert of Little Rann of Kutch, India: A Phylogenetic Perspective

Efficiency of Microencapsulation of Proteolytic Enzymes

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