Production and biochemical characterization of an alkaline protease from Aspergillus oryzae CH93

Salihi, Ahsan; Asoodeh, Ahmad; Aliabadian, Mansour

doi:10.1016/j.ijbiomac.2016.06.023

Cited by 43 publications

(25 citation statements)

References 40 publications

(24 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The total activity and total protein of the crude enzyme showed a notably high specific activity of 439062U and 1934.7 mg, which is markedly higher than the enzyme activities previously reported in A. oryzae CH93 (14.7 U/g) (Salihi et al 2016 ) and Aeromonas veronii PG01 (18.5 U) (Divakar et al 2010 ). The crude enzyme was initially concentrated from 20 to 90% using ammonium sulfate.…”

Section: Resultsmentioning

confidence: 53%

Purification and characterization of neutral protease from Aspergillus oryzae Y1 isolated from naturally fermented broad beans

et al. 2018

AMB Expr

View full text Add to dashboard Cite

The strain Y1, with a notably high production of neutral protease, was isolated from naturally fermented broad beans and subsequently identified as Aspergillus oryzae, through the analysis of its morphology characteristics and 18S rDNA sequence. Naturally fermented broad beans are the main raw material in Sichuan broad-bean sauce. The neutral protease from Aspergillus oryzae Y1 was purified using ammonium sulphate precipitation and DEAE-Sepharose Fast Flow chromatography, which resulted in a 10.0-fold increase in the specific activity (2264.3 U/mg) and a recovery rate of 21%. The estimated molecular mass of the purified protease was approximately 45 kDa. The optimal pH and temperature of the purified protease were 7.0 and 55 °C, respectively. The heat resistance of the purified protease was significantly higher than the commercial protease. The effect of metal ions on the activity of the purified protease approximated that of commercial neutral protease. Furthermore, the maximum hydrolysis rate (Vmax) and apparent Michaelis–Menten constant (Km) values of the purified protease were 256.4103 μg/mL min and 20.0769 mg/mL, respectively. The purified protease had a higher affinity for the substrate than the commercial neutral protease. All the results suggest that this neutral protease exhibits the potential for application in industry due to its good resistance to high temperatures and wide range of acids and bases.

show abstract

Section: Resultsmentioning

confidence: 53%

Purification and characterization of neutral protease from Aspergillus oryzae Y1 isolated from naturally fermented broad beans

et al. 2018

AMB Expr

View full text Add to dashboard Cite

show abstract

“…AprA exhibits higher optimal pH (10.5) and better stability under alkaline conditions than many previously reported alkaline proteases from Bacillus 4 , 6 , 8 , 9 , 15 , 16 , 27 , 33 , 51 , 56 – 61 , Vibrio 19 , Aspergillus 21 , 62 , 63 , Thermoactinomyces 24 , 26 , Streptomyces 25 , Thermus 28 , Pseudoalteromonas 41 , Alteromonas 42 , Stenotrophomonas 44 , Trametes 52 , Termitomyces 54 , Virgibacillus 64 , Caldicoprobacter 65 , Hirsutella 66 , Scopulariopsis 67 , and Penicillium 68 (Table S1 ). Furthermore, AprA also retains high activity and stability over a wide range of pH (7.0–11.5) and temperature (40–70 °C), several metal ions, surfactants and some oxidizing and reducing agents.…”

Section: Discussionmentioning

confidence: 82%

“…In addition to Bacillus bacterial, alkaline proteases from several gram-negative bacteria such as Pseudomonas 17 , Xanthomonas 18 , and Vibrio 19 have also been found. Moreover, fungi especially genus Aspergillus 20 , 21 , yeast 22 , 23 , and actinomycetes 24 , 25 are also prolific sources for alkaline proteases. Most of these alkaline proteases are either serine proteases or metalloproteases 3 , and show optimal pH around 8–12 and optimal temperature around 40–65 °C.…”

Section: Introductionmentioning

confidence: 99%

A novel alkaline protease from alkaliphilic Idiomarina sp. C9-1 with potential application for eco-friendly enzymatic dehairing in the leather industry

Zhou

Qin

Chen

et al. 2018

Sci Rep

View full text Add to dashboard Cite

Alkaline proteases have a myriad of potential applications in many industrial processes such as detergent, food and feed production, waste management and the leather industry. In this study, we isolated several alkaline protease producing bacteria from soda lake soil samples. A novel serine alkaline protease (AprA) gene from alkaliphilic Idiomarina sp. C9-1 was cloned and expressed in Escherichia coli. The purified AprA and its pre-peptidase C-terminal (PPC) domain-truncated enzyme (AprA-PPC) showed maximum activity at pH 10.5 and 60 °C, and were active and stable in a wide range of pH and temperature. Ca2+ significantly improved the thermostability and increased the optimal temperature to 70 °C. Furthermore, both AprA and AprA-PPC showed good tolerance to surfactants and oxidizing and reducing agents. We found that the PPC domain contributed to AprA activity, thermostability and surfactant tolerance. With casein as substrate, AprA and AprA-PPC showed the highest specific activity of 42567.1 U mg−1 and 99511.9 U mg−1, the Km values of 3.76 mg ml−1 and 3.98 mg ml−1, and the Vmax values of 57538.5 U mg−1 and 108722.1 U mg−1, respectively. Secreted expression of AprA-PPC in Bacillus subtilis after 48 h cultivation resulted in yield of 4935.5 U ml−1 with productivity of 102.8 U ml−1 h−1, which is the highest reported in literature to date. Without adding any lime or sodium sulfide, both of which are harmful pollutants, AprA-PPC was effective in dehairing cattle hide and skins of goat, pig and rabbit in 8–12 h without causing significant damage to hairs and grain surface. Our results suggest that AprA-PPC may have great potentials for ecofriendly dehairing of animal skins in the leather industry.

show abstract

“…Unfortunately, the current quality of soy sauce and the utilization rate of raw materials in China are still low, leading to their weak competitiveness in both foreign and domestic markets . APs from other strains, such as Bacterium sp., Bacillus subtilis , Streptomyces clavuligerus and A. oryzae , were found to be salt‐tolerant. However, to date, comparatively little has been revealed about the biochemical properties and salt‐tolerant mechanisms of AP produced from A. oryzae 3.042.…”

Section: Introductionmentioning

confidence: 99%

Separation, biochemical characterization and salt‐tolerant mechanisms of alkaline protease from Aspergillus oryzae

et al. 2019

View full text Add to dashboard Cite

BACKGROUND The salt tolerance of proteases secreted by Aspergillus oryzae 3.042 closely relates to the utilization of raw materials and the quality of soy sauce. However, little is known about the salt‐tolerant proteases and their salt‐tolerant mechanisms. RESULTS In this study, we isolated and identified a salt‐tolerant alkaline protease (AP, approximately 29 kDa) produced by A. oryzae 3.042. It was considered as a metal‐ion‐independent serine protease. The optimum and stable pH values were both pH 9.0 and the optimum temperature was 40 °C. Over 20% relative activity of AP remained in the presence of 3.0 mol L−1 NaCl after 7 days, but its Km and Vmax were only mildly influenced by the presence of 3.0 mol L−1 NaCl, indicating its outstanding salt tolerance. Furthermore, AP was more stable than non‐salt‐tolerant protease at high salinity. The salt‐tolerant mechanisms of AP could be due to more salt bridges, higher proportion of ordered secondary structures and stronger hydrophobic amino acid residues in the interior. CONCLUSIONS The above results are vital for maintaining, activating and/or modulating the activity of AP in high‐salt environments. They would also provide theoretical guidance for the modification of AP and the engineering of A. oryzae 3.042 so as to secrete more AP. © 2018 Society of Chemical Industry

show abstract

Production and biochemical characterization of an alkaline protease from Aspergillus oryzae CH93

Cited by 43 publications

References 40 publications

Purification and characterization of neutral protease from Aspergillus oryzae Y1 isolated from naturally fermented broad beans

Purification and characterization of neutral protease from Aspergillus oryzae Y1 isolated from naturally fermented broad beans

A novel alkaline protease from alkaliphilic Idiomarina sp. C9-1 with potential application for eco-friendly enzymatic dehairing in the leather industry

Separation, biochemical characterization and salt‐tolerant mechanisms of alkaline protease from Aspergillus oryzae

Contact Info

Product

Resources

About