Characterization of a New Intracellular Alginate Lyase with Metal Ions-Tolerant and pH-Stable Properties

BACKGROUND: Alginate oligosaccharides (AOS), with various physiological effects, have been widely used in the food, agricultural, and pharmaceutical industries. The biological enzymatic method of preparing AOS, using alginate lyase, has more advantages compared with physical and chemical methods. Cloning and heterologously expressing alginate lyase are therefore very important.RESULTS: A novel alginate lyase, BY17PV7, from Microbulbifer sp. BY17, isolated from Gracilaria, was cloned and expressed in Escherichia coli BL21(DE3). BY17PV7 was about 27 KDa. BY17PV7 showed the greatest activity (150.42 ± 3.32 U/mg) at 43 °C and pH 8.9. It could be activated by Ca 2+ , Mn 2+ , Co 2+ , Fe 3+ , Na + , and inhibited by Mg 2+ , Zn 2+ , Ba 2+ , Cu 2+ , sodium dodecyl sulfate (SDS), ethylene diamine tetraacetic acid (EDTA). BY17PV7 had a wide range of substrate specificity and good degradation effects for poly ⊎-D-mannuronate (polyM) and poly ⊍-L-guluronate (polyG), demonstrating that it is a bifunctional alginate lyase. The kinetic parameters showed that BY17PV7 had a greater affinity for polyG. BY17PV7 released AOS with a degree of polymerization (DP) of 3-4 in an endolytic manner from sodium alginate. Alginate oligosaccharides showed strong antioxidant ability of reducing Fe 3+ and scavenging radicals such as hydroxyl, 2,2-azion-bia (3-ethylbenzo-thiazoline-6-sulfonic acid diammonium salt) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH).CONCLUSION: A novel bifunctional alginate lyase, BY17PV7, was expressed and characterized in Escherichia coli BL21(DE3). The results were helpful for the analysis of the molecular mechanisms of degrading patterns in the polysaccharide lyase (PL) family.

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“…W2. [42][43][44] Similarly, BY17PV7 preferred to degrade polyG according to substrate specificity (Fig. 4(G)).…”

Section: Resultsmentioning

confidence: 89%

“…Similarly, A1m (PL7) from Agarivorans sp. displayed maximum enzyme activity at pH 10.0 and was maintained in a stable way in the range of 6.0–8.0, 50 and Alyw202 showed the maximum enzyme at pH 9.0 and remained stable in the range of pH 3.0–10.0 42 …”

Section: Resultsmentioning

confidence: 98%

Cloning, expression, and characterization of a novel endo‐type alginate lyase from Microbulbifer sp. BY17

et al. 2022

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“…For example, Aly08 from PL7 can remain more than 80% of its initial activity in a wide pH range (4.0-10.0), and its optimal pH was found to be 8.35 [64]. In particular, it can retain more than 60% of initial activity in a wide pH range from 7.0 to 11.0 after incubation in different buffers at 4 • C for 12 h. Similar to Aly08, Alyw202 from PL7 exhibited the highest activity at pH 9.0, and it showed a wider pH range than Aly08 [66]. Alginate lyases with excellent pH stability and a wide pH range could be candidates for hydrolyzing acid or alkaline pretreated alginate, which are generally needed to produce alginate oligosaccharides [16].…”

Section: Wide Ph Adaptation Range and Alkaline Propertymentioning

confidence: 92%

Structure Characteristics, Biochemical Properties, and Pharmaceutical Applications of Alginate Lyases

et al. 2021

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Alginate, the most abundant polysaccharides of brown algae, consists of various proportions of uronic acid epimers α-L-guluronic acid (G) and β-D-mannuronic acid (M). Alginate oligosaccharides (AOs), the degradation products of alginates, exhibit excellent bioactivities and a great potential for broad applications in pharmaceutical fields. Alginate lyases can degrade alginate to functional AOs with unsaturated bonds or monosaccharides, which can facilitate the biorefinery of brown algae. On account of the increasing applications of AOs and biorefinery of brown algae, there is a scientific need to explore the important aspects of alginate lyase, such as catalytic mechanism, structure, and property. This review covers fundamental aspects and recent developments in basic information, structural characteristics, the structure–substrate specificity or catalytic efficiency relationship, property, molecular modification, and applications. To meet the needs of biorefinery systems of a broad array of biochemical products, alginate lyases with special properties, such as salt-activated, wide pH adaptation range, and cold adaptation are outlined. Withal, various challenges in alginate lyase research are traced out, and future directions, specifically on the molecular biology part of alginate lyases, are delineated to further widen the horizon of these exceptional alginate lyases.

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“…NAP1 (AlgC-PL7) [ 107 ], Streptomyces luridiscabiei (AlyDS44) [ 100 ], Vibrio sp. W2 (Alyw202 and Alyw203) [ 105 , 119 ], which had the optimum working temperature of 45 °C. The optimal temperature for Aly1281 from Pseudoalteromonas carrageenovora ASY5 [ 108 ], AlgMsp from Microbulbifer sp.…”

Section: Biochemical Properties Of Marine Bacteria-produced Alginate ...mentioning

confidence: 99%

Alginate Lyases from Marine Bacteria: An Enzyme Ocean for Sustainable Future

et al. 2022

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The cell wall of brown algae contains alginate as a major constituent. This anionic polymer is a composite of β-d-mannuronate (M) and α-l-guluronate (G). Alginate can be degraded into oligosaccharides; both the polymer and its products exhibit antioxidative, antimicrobial, and immunomodulatory activities and, hence, find many commercial applications. Alginate is attacked by various enzymes, collectively termed alginate lyases, that degrade glycosidic bonds through β-elimination. Considering the abundance of brown algae in marine ecosystems, alginate is an important source of nutrients for marine organisms, and therefore, alginate lyases play a significant role in marine carbon recycling. Various marine microorganisms, particularly those that thrive in association with brown algae, have been reported as producers of alginate lyases. Conceivably, the marine-derived alginate lyases demonstrate salt tolerance, and many are activated in the presence of salts and, therefore, find applications in the food industry. Therefore, this review summarizes the structural and biochemical features of marine bacterial alginate lyases along with their applications. This comprehensive information can aid in the expansion of future prospects of alginate lyases.

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Characterization of a New Intracellular Alginate Lyase with Metal Ions-Tolerant and pH-Stable Properties

Cited by 17 publications

References 41 publications

Cloning, expression, and characterization of a novel endo‐type alginate lyase from Microbulbifer sp. BY17

Cloning, expression, and characterization of a novel endo‐type alginate lyase from Microbulbifer sp. BY17

Structure Characteristics, Biochemical Properties, and Pharmaceutical Applications of Alginate Lyases

Alginate Lyases from Marine Bacteria: An Enzyme Ocean for Sustainable Future

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