Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications

Slavić, Marinela Šokarda; Kojić, Milan; Margetić, Aleksandra; Stanisavljević, Nemanja; Gardijan, Lazar; Božić, Nataša; Vujčić, Zoran

doi:10.1016/j.ijbiomac.2023.126055

Cited by 3 publications

(5 citation statements)

References 61 publications

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“…This preference may be attributed to the relatively lower amylose content of wheat starch (24%) compared with the other starch sources (27% on average) (Table 3 ). In general, starches with low amylose contents (e.g., wheat) have less compact granule structures, providing better accessibility and higher freedom for the enzyme to react with the starches, resulting in greater degradation efficiency, as supported by earlier reports (Božić et al 2011 ; Chakraborty et al 2020 ; Slavić et al 2023 ; Tester et al 2004 ; Wang et al 2022 ). In relative terms, starches with high amylose contents (e.g., tapioca, sago, potato, rice, and corn) have densely packed granule structures, which limits enzyme access and makes them less prone to enzymatic action (Bertoft 2017 ; Chakraborty et al 2020 ; Wang et al 2022 ).…”

Section: Discussionmentioning

confidence: 67%

“…In contrast, thermophile enzymes (e.g., Anoxybacillus and Geobacillus spp.) have higher optimum temperatures, ranging from 65 to 105 °C (Diderichsen et al 1991 ; Mehta and Satyanarayana 2013 ; Slavić et al 2023 ; Tawil et al 2012 ). Regarding the optimal pH, most GH13_5 α-amylases displayed optimal activity under acidic conditions, within the pH range of 4.0–6.9 (Table 1 ).…”

Section: Discussionmentioning

confidence: 99%

“…This positive influence of calcium chloride has been observed in other GH13_5 bacterial α-amylases, such as those from Pontibacillus sp. ZY, P. mobilis , and A. vranjensis ST4 (Fang et al 2019 ; Jabbour et al 2013 ; Slavić et al 2023 ). In the homology model of AmyJM, a predicted calcium-binding site was identified to harbor a Ca 2+ ion (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…Using raw starch-degrading α-amylases could reduce energy consumption by approximately 15% compared with traditional physical or chemical processes (Sun et al 2010 ). Therefore, raw starch-degrading α-amylases have potential industrial application as a non-thermal processing strategy to the traditional heating process in starch liquefaction (Božić et al 2020 ; Fang et al 2019 ; Slavić et al 2023 ).…”

Section: Introductionmentioning

confidence: 99%

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Exploring a novel GH13_5 α-amylase from Jeotgalibacillus malaysiensis D5T for raw starch hydrolysis

Radzlin,

Mohamad Ali,

Goh

et al. 2024

AMB Expr

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Abstractα-Amylase plays a crucial role in the industrial degradation of starch. The genus Jeotgalibacillus of the underexplored marine bacteria family Caryophanaceae has not been investigated in terms of α-amylase production. Herein, we report the comprehensive analysis of an α-amylase (AmyJM) from Jeotgalibacillus malaysiensis D5T (= DSM28777T = KCTC33550T). Protein phylogenetic analysis indicated that AmyJM belongs to glycoside hydrolase family 13 subfamily 5 (GH13_5) and exhibits low sequence identity with known α-amylases, with its closest counterpart being the GH13_5 α-amylase from Bacillus sp. KSM-K38 (51.05% identity). Purified AmyJM (molecular mass of 70 kDa) is stable at a pH range of 5.5–9.0 and optimally active at pH 7.5. The optimum temperature for AmyJM is 40 °C, where the enzyme is reasonably stable at this temperature. Similar to other α-amylases, the presence of CaCl2 enhanced both the activity and stability of AmyJM. AmyJM exhibited activity toward raw and gelatinized forms of starches and related α-glucans, generating a mixture of reducing sugars, such as glucose, maltose, maltotriose, maltotetraose, and maltopentaose. In raw starch hydrolysis, AmyJM exhibited its highest efficiency (51.10% degradation) in hydrolyzing raw wheat starch after 3-h incubation at 40 °C. Under the same conditions, AmyJM also hydrolyzed tapioca, sago, potato, rice, and corn raw starches, yielding 16.01–30.05%. These findings highlight the potential of AmyJM as a biocatalyst for the saccharification of raw starches, particularly those derived from wheat.

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Section: Discussionmentioning

confidence: 67%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Exploring a novel GH13_5 α-amylase from Jeotgalibacillus malaysiensis D5T for raw starch hydrolysis

Radzlin,

Mohamad Ali,

Goh

et al. 2024

AMB Expr

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“…However, the room temperature amylases exhibit poor thermal stability when exposed to high temperature conditions, resulting in a rapid decline in enzymatic activity (Gupta et al, 2003). Geobacillus stearothermophilus α-amylase, derived from a thermophilic sporeforming bacterium, belongs to a class of starch enzymes with significant potential for industrial applications (Srivastava and Baruah, 1986;Offen et al, 2015;Slavić et al, 2023). However, in modern starch processing processes, such as ethanol production, amylases are required to have higher temperature stability (the reaction temperatures typically exceeding 90°C).…”

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confidence: 99%

Modifying the amino acids in conformational motion pathway of the α-amylase of Geobacillus stearothermophilus improved its activity and stability

Hu,

Hong,

et al. 2023

Front. Microbiol.

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Amino acids along the conformational motion pathway of the enzyme molecule correlated to its flexibility and rigidity. To enhance the enzyme activity and thermal stability, the motion pathway of Geobacillus stearothermophilus α-amylase has been identified and molecularly modified by using the neural relational inference model and deep learning tool. The significant differences in substrate specificity, enzymatic kinetics, optimal temperature, and thermal stability were observed among the mutants with modified amino acids along the pathway. Mutants especially the P44E demonstrated enhanced hydrolytic activity and catalytic efficiency (kcat/KM) than the wild-type enzyme to 95.0% and 93.8% respectively, with the optimum temperature increased to 90°C. This mutation from proline to glutamic acid has increased the number and the radius of the bottleneck of the channels, which might facilitate transporting large starch substrates into the enzyme. The mutation could also optimize the hydrogen bonding network of the catalytic center, and diminish the spatial hindering to the substrate entry and exit from the catalytic center.

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Thermostable α-Amylases and Laccases: Paving the Way for Sustainable Industrial Applications

Jaiswal,

Jaiswal

2024

Processes

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The growing demand in industrial and biotechnological settings for more efficient enzymes with enhanced biochemical features, particularly thermostability and thermotolerance, necessitates a timely response. Renowned for their versatility, thermostable enzymes offer significant promise across a range of applications, including agricultural, medicinal, and biotechnological domains. This comprehensive review summarizes the structural attributes, catalytic mechanisms, and connection between structural configuration and functional activity of two major classes of thermostable enzymes: α-amylases and laccases. These enzymes serve as valuable models for understanding the structural foundation behind the thermostability of proteins. By highlighting the commercial importance of thermostable enzymes and the interest these generate among researchers in further optimization and innovation, this article can greatly contribute to ongoing research on thermostable enzymes and aiding industries in optimizing production processes via immobilization, use of stabilizing additives, chemical modification, protein engineering (directed evolution and mutagenesis), and genetic engineering (through cloning and expression of thermostable genes). It also gives insights to the exploration of suitable strategies and factors for enhancing thermostability like increasing substrate affinity; introducing electrostatic, intramolecular, and intermolecular hydrophobic interactions; mitigating steric hindrance; increasing flexibility of an active site; and N- and C-terminal engineering, thus resulting in heightened multipronged stability and notable enhancements in the enzymes’ industrial applicability.

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Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications

Cited by 3 publications

References 61 publications

Exploring a novel GH13_5 α-amylase from Jeotgalibacillus malaysiensis D5T for raw starch hydrolysis

Exploring a novel GH13_5 α-amylase from Jeotgalibacillus malaysiensis D5T for raw starch hydrolysis

Modifying the amino acids in conformational motion pathway of the α-amylase of Geobacillus stearothermophilus improved its activity and stability

Thermostable α-Amylases and Laccases: Paving the Way for Sustainable Industrial Applications

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