Purification and characterization of extracellular glutaminase from Aspergillus oryzae NRRL 32567

Bazaraa, W. A.; Alian, A.; El-Shimi, Nagwa M.; Mohamed, Reda Mahgoub

doi:10.1016/j.bcab.2016.02.009

Cited by 12 publications

(3 citation statements)

References 28 publications

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“…thermodenitrificans, respectively [15,42,43] (Supplementary Table S1). Compared to mesophilic counterparts, the L-glutamine glutaminase from Aspergillus oryzae, whose optimum temperature is 30 °C, retained only 40% of maximum activity at 25 °C [47], whereas the L-glutamine glutaminase from E. coli showed low level of activity at 4 °C [48].…”

Section: Effect Of Temperature On Enzyme Activity and Stabilitymentioning

confidence: 92%

Characteristics of a Cold-Adapted L-glutaminase with Potential Applications in the Food Industry

Ferreira

Herrmann-Andrade

Binolfi

et al. 2021

Appl Biochem Biotechnol

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L-glutaminases are enzymes that catalyze the hydrolysis of L-glutamine, producing L-glutamate and ammonium, and they have promising applications in pharmaceutical and food industries. Several investigations have focused on thermotolerant L-glutaminases; however, studies on cold-adapted L-glutaminases have not been reported. These enzymes could be useful in the food industry because they display high catalytic activity at low and room temperatures, a valuable feature in processes aimed to save energy. Besides, they can be easily inactivated by warming and are suitable to prevent decomposition of thermo-labile compounds. The objectives of this work were to characterize the L-glutaminase from the Antarctic bacterium Bizionia argentinensis and analyze its capability as flavor enhancer of protein hydrolysates. The enzyme was heterologously expressed and purified from Escherichia coli, obtaining optimum and homogeneous yields. Kinetic parameters K m and V max were located at the lower and upper range of values reported for L-glutaminases, suggesting high catalytic efficiency. Optimum temperature was 25 °C, and the enzyme conserved around 90% of maximum activity at 0 °C and in presence of 15% (v/v) ethanol and methanol. In saline conditions, the enzyme conserved around 80% of maximum activity in 3 M NaCl. Analysis of structural model suggested cold-adaptation features such as low Arg/ (Arg+Lys) ratio and fewer intramolecular interactions than mesophilic and thermo-tolerant L-glutaminases. This work provides a novel cold-adapted L-glutaminase with promising features in the food industry.

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Section: Effect Of Temperature On Enzyme Activity and Stabilitymentioning

confidence: 92%

Characteristics of a Cold-Adapted L-glutaminase with Potential Applications in the Food Industry

Ferreira

Herrmann-Andrade

Binolfi

et al. 2021

Appl Biochem Biotechnol

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“…Watanabe et al, (2016) [12] observed that glucosidase thermostability maintained residual activity at almost 100% when incubated at 4 ° C and in the range 20-60°C, losing all of its residual activity between 70-80°C. These results were obtained when studying the purified glucosidase obtained from Aspergillus oryzae at temperatures of 4-80°C in phosphate buffer (pH 6.8).…”

Section: Stability In Temperature Differentmentioning

confidence: 99%

Stability study of pre-purified protease obtained from Aspergillus oryzae NRRL 1911 by solid state fermentation with canola cake as substrate at different pH and temperature

Souza

Nogueira

Dutra

et al. 2019

BJD

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“…2), an enzyme that can produce glutamic acid from glutamine, which is plentiful in the fermentation broth ( Binod et al, 2017 ). To be noted, a high salt concentration was required during soy sauce fermentation by Aspergillus oryzae Ye et al (2013) , and 15–18% (w/v) of sodium chloride is typically used ( Koibuchi et al, 2000 ; Ito et al, 2013 ; Bazaraa et al, 2016 ). However, the catalytic activity of A. oryzae glutaminase was significantly inhibited at NaCl concentrations of over 15% ( Masuo et al, 2005 ).…”

Section: Introductionmentioning

confidence: 99%

Enhanced salt-tolerance of Bacillus subtilis glutaminase by fusing self-assembling amphipathic peptides at its N-terminus

Liu

Rao

Chen

et al. 2022

Front. Bioeng. Biotechnol.

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Glutaminase (EC 3.5.1.2) can catalyze the deamidation of glutamine, which has been used to improve umami taste in oriental fermented foods. However, a high salt concentration is still a fundamental challenge for glutaminase application, especially in soy sauce production. To improve the salt tolerance of glutaminase, the self-assembling amphiphilic peptides EAK16 and ELK16 were fused to the N-terminus of a mutant (E3C/E55F/D213T) derived from Bacillus subtilis glutaminase, yielding the fusion enzymes EAK16-E3C/E55F/D213T and ELK16-E3C/E55F/D213T, respectively. As ELK16-E3C/E55F/D213T was expressed as insoluble active inclusion bodies, only the purified EAK16-E3C/E55F/D213T was subjected to further analyses. After the incubation with 18% (w/v) NaCl for 200 min, the residual activities of EAK16-E3C/E55F/D213T in a NaCl-free solution reached 43.6%, while E3C/E55F/D213T was completely inactivated. When the enzyme reaction was conducted in the presence of 20% NaCl, the relative activity of EAK16-E3C/E55F/D213T was 0.47-fold higher than that of E3C/E55F/D213T. As protein surface hydrophobicity and protein particle size analysis suggested, oligomerization may play an important role in the salt-tolerance enhancement of the fusions. Furthermore, EAK16-E3C/E55F/D213T achieved a 0.88-fold increase in the titer of glutamic acid in a model system of soy sauce fermentation compared to E3C/E55F/D213T. Therefore, the fusion with self-assembling amphiphilic peptides is an efficient strategy to improve the salt-tolerance of glutaminase.

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Purification and characterization of extracellular glutaminase from Aspergillus oryzae NRRL 32567

Cited by 12 publications

References 28 publications

Characteristics of a Cold-Adapted L-glutaminase with Potential Applications in the Food Industry

Characteristics of a Cold-Adapted L-glutaminase with Potential Applications in the Food Industry

Stability study of pre-purified protease obtained from Aspergillus oryzae NRRL 1911 by solid state fermentation with canola cake as substrate at different pH and temperature

Enhanced salt-tolerance of Bacillus subtilis glutaminase by fusing self-assembling amphipathic peptides at its N-terminus

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