Screening of Thermostable Protease Producing Microorganisms Isolated From Indonesian Hotspring

The gut wastes of Sardinella longiceps were used as substrate for protease production. The gut waste has 61.6% proteins, 21.8% lipids, 8.5% carbohydrates on dry weight basis and trace elements. The significant factors of protease fermentation were screened by Plackett-Burman design. A protease activity of 68.56 U/ml was predicted at 46.31 °C, incubation time 71.11 h, inoculum 4.86% (v/v) and substrate concentration 2.66% (w/v), using response surface methodology. However, the validation experiment showed 73.52 U/ml activity. The artificial neural network was found as a better tool to predict the experimental results. The partially purified protease showed higher activity at pH 9 and 10 and retained 90% activity after 120 h at pH 9. It showed maximum activity at 50 °C and retained 88% residual activity until 90 min at 50 °C. Zn++ enhanced the protease activity by 40%. The protease retained an activity of 93, 103, 90 and 98% against urea, β-mercaptoethanol, SDS and tween 80 respectively. The alkaline protease was compatible with all the commercial detergents tested with the residual activity above 90%. The alkaline protease exhibited 22% higher activity on the tryptone soya substrate. The gut waste of S. longiceps is a worthy low cost substrate for the production of industrially important alkaline protease.

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“…and Brevibacillus spp. isolated from various hot springs across the world were reported to produce thermostable proteases 9,10 .…”

Section: Resultsmentioning

confidence: 99%

Production of thermotolerant, detergent stable alkaline protease using the gut waste of Sardinella longiceps as a substrate: Optimization and characterization

Ramkumar

Sivakumar

Gujarathi

et al. 2018

Sci Rep

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“…Lampung is known as hot spring spots, for instane, in Kalianda, Padang Cermin (Zilda et al, 2012) and Wartawan beach. The presence of hot springs is mainly associated with volcano activity in the area (Kruse, 1999).…”

Section: The Environmental Conditions Of Sampling Locationmentioning

confidence: 99%

“…The hot environments are spread in many locations, not only in the land but also coastal environments including deep sea area. Some of these areas are located at Tambrana, Central Sulawesi (Zilda, Kusumarini, & Chasanah, 2008), Poso, Central Sulawesi (Zilda, Patantis, & Chasanah, 2009), Padang Cermin and Kalianda, Lampung and Banyu Wedang, Bali (Zilda et al, 2012). These hot environments are characterized by a mat layer, made up from microbial mat communities.…”

Section: Introductionmentioning

confidence: 99%

Bacterial Diversity of a Microbial Mat from Hot Spring at Wartawan Beach, Lampung and Its Potential as a Source of Hydrogenases

Patantis

Chasanah

Fawzya³

et al. 2018

Squalen Bull. Marine Fisheries Postharvest Biotech.

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Biohydrogen produced from thermophilic hydrogenases is an ideal and clean energy sources. As the biggest tectonic area in the world, Indonesia is potential for thermophile isolation. The aims of this study were to analyze the bacterial diversity of a microbial mat from hot spring at Wartawan beach, Lampung and to analyze the potency of microbial mat for hydrogenases, using clone library method. The diversity of 16S rRNA showed that the microbial mat sample contained 9 phyla of bacteria, and dominated by Cyanobacteria and Proteobacteria. These p hyla indicate that the bacterial community of the microbial mat consisted of phototrophic and heterotrophic groups. In addition, a microbial mat of W artawan beach environment might be influenced by marine environment an d hydrothermal vent which was indicated by detection of both associated bacteria. The diversity of hydrogenase genes using NiFe hydrogenase (NiFe) and FeFe hydrogenase (FeFe) genes showed that Cyanobacteria was specifically related to NiFe, while Firmicutes was associated with FeFe. Proteobacteria and Bacteroidetes, however, were detected for both genes. The detected hydrogenase genes indicate that the microbial mat from hot spring at W artawan beach is a promising source for hydrogenases isolation and further applications for biohydrogen production as a renewable energy.

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“…Hydrolase production by thermotolerant bacterial isolates was screened by plating on starch (Khalil, 2011), carboxymethyl cellulose (Teather and Wood, 1982), skimmed milk (Zilda et al, 2012), pectin (Huang et al, 2012) and xylan (Gessesse and Gashe, 1997) agar plates for amylase, cellulose, protease, pectinase and xylanase activity respectively. All the assays were conducted at 55°C for 48 h. …”

Section: Detection Of Enzymesmentioning

confidence: 99%

Thermotolerant bacteria of biotechnological potential from hot springs in Eritrea

Ghilamicael¹,

Budambula²,

Anami³

et al. 2018

Afr. J. Microbiol. Res.

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Thermophiles are excellent sources of enzymes that can withstand and carry out reactions efficiently under high temperatures. This study isolated and characterised thermotolerant bacteria that produce enzymes of potential industrial value from five hot springs in Eritrea. A total of 65 bacterial isolates were obtained from the five hot springs. Out of the 65 isolates; 19 isolates produced a positive reaction for amylases, 36 for carboxymethyl cellulases, eight for proteases, 10 for xylanases and 11 for pectinases. More than half (36 out of 65) were able to produce carboxymethyl cellulases. Six isolates which showed carboxymethyl cellulase activity were from the genus Bacillus, while those belonging to Brevibacillus were seven. BLAST analysis of the partial sequences showed that 19 out of the 24 isolates sequenced showed high similarity (> 99%) to those of reference strains of the genera Bacillus and Brevibacillus available in the Genebank and EZ-taxon databases. The five isolates (E5, G2, G8, M1 and M13) that showed moderate similarities (97.2-99%) to strains from the Genebank and EZ-taxon databases were further characterized. Physiological characterization of the five selected isolates based on tolerance to NaCl, temperature and production of hydrolytic enzymes indicated that these isolates are potentially novel. Isolates G8 and M13 showed significantly higher amylase activity (p < 0.05) than the other three isolates. Caseinase activity recorded by the five isolates was the highest (p < 0.05) compared to other enzyme activities. The enzymes produced by thermotolerant bacteria from the five hot springs may be potentially useful for catalysis under harsh operational conditions encountered in industrial processes.

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Screening of Thermostable Protease Producing Microorganisms Isolated From Indonesian Hotspring

Cited by 23 publications

References 23 publications

Production of thermotolerant, detergent stable alkaline protease using the gut waste of Sardinella longiceps as a substrate: Optimization and characterization

Production of thermotolerant, detergent stable alkaline protease using the gut waste of Sardinella longiceps as a substrate: Optimization and characterization

Bacterial Diversity of a Microbial Mat from Hot Spring at Wartawan Beach, Lampung and Its Potential as a Source of Hydrogenases

Thermotolerant bacteria of biotechnological potential from hot springs in Eritrea

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