Construction and biochemical characterization of a novel hybrid alginate lyase with high activity by module recombination to prepare alginate oligosaccharides

Hu, Fu; Cao, Shengsheng; Li, Qian; Zhu, Benwei; Yao, Zhong

doi:10.1016/j.ijbiomac.2020.11.009

Cited by 18 publications

(5 citation statements)

References 30 publications

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“…Compared to the previous studies, most of the reported enzymes were basophilic lyases and showed pH stability at higher salt concentrations (pH 8.5) [ 22 ]. Earlier studies have reported that the optimum activity of alginate lyase was at acidic pH 6 [ 23 ] and basic pH of 8 to 10 [ 24 , 25 , 26 , 27 , 28 ].…”

Section: Resultsmentioning

confidence: 99%

Production of Alginate Oligosaccharides (AOSs) Using Enhanced Physicochemical Properties of Immobilized Alginate Lyase for Industrial Application

Kaur,

Abraham,

Franco

et al. 2024

Marine Drugs

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Alginate lyase (AL) is a polysaccharide-degrading enzyme that can degrade alginate by hydrolyzing glycosidic bonds and produces unsaturated alginate oligosaccharides (AOSs). These AOSs have wide therapeutic and nutraceutical applications. However, to produce alginate oligosaccharides in a cost-effective manner is challenging due to the low availability and high cost of this degrading enzyme. Immobilization of the enzyme facilitates industrial applications owing to its stability, reusability, and cost-effectiveness. This study was focused on the enhancement of the properties of alginate lyase and improvement of the production of AOS. Alginate lyase was immobilized on magnetic nanoparticles (NPs) using glutaraldehyde as the crosslinker. The study showed that the maximum binding achieved between NPs and protein in the enzyme was 71% at a ratio of 1:150 NP:protein. As a result of immobilization, the optimum activity of free enzyme which was obtained at 37 °C and pH 7.4 changed to 45 °C and pH 9. Furthermore, the enzyme was thermostable at 45 °C for 3 h with up to 50% reusability for six consecutive cycles. Storage stability after 15 days showed ~67% relative hydrolysis of alginate. The free alginate lyase (25 IU) showed 76% raw biomass (seaweed) hydrolysis which is higher compared to 63% provided by the immobilized enzyme. As a result of efficient hydrolysis, AOSs with molecular weight profile of 370–1040 kDa were produced and detected using HPLC.

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Section: Resultsmentioning

confidence: 99%

Production of Alginate Oligosaccharides (AOSs) Using Enhanced Physicochemical Properties of Immobilized Alginate Lyase for Industrial Application

Kaur,

Abraham,

Franco

et al. 2024

Marine Drugs

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“…To measure the effects of other metal ions, the EDTA-pretreated purified enzymes were incubated with 1 mM of various metal ions (Fe 3+ , Fe 2+ , Co 2+ , Ni 2+ , Cu 2+ , Zn 2+ , Ca 2+ , Mg 2+ , Mn 2+ , Na + , and K + ) at 4 °C overnight. The activity of the control was defined as a relative activity of 100% [ 35 ].…”

Section: Methodsmentioning

confidence: 99%

Characterization of a Novel Polysaccharide Lyase Family 5 Alginate Lyase with PolyM Substrate Specificity

Zhou

Meng

Zhang

et al. 2022

Foods

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Alginate lyases (ALyases) have been widely applied in enzymatically degrading alginate for the preparation of alginate oligosaccharides (AOS), which possess a range of excellent physiological benefits including immunoregulatory, antivirus, and antidiabetic properties. Among the characterized ALyases, the number of ALyases with strict substrate specificity which possess potential in directed preparation of AOS is quite small. ALyases of polysaccharides lyase (PL) 5 family have been reported to perform poly-β-D-mannuronic acid (Poly-M) substrate specificity. However, there have been fewer studies with a comprehensive characterization and comparison of PL 5 family ALyases. In this study, a putative PL 5 family ALyase PMD was cloned from Pseudomonas mendocina and expressed in Escherichia coli. The novel ALyase presented maximum activity at 30 °C and pH 7.0. PMD displayed pH stability properties under the range of pH 5 to pH 9, which retained more than 80% relative activity, even when incubated for 48 h. Product analysis indicated that PMD might be an endolytic ALyase with strict Poly M substrate specificity and yield disaccharide and trisaccharide as main products. In addition, residues K58, R66, Y248, and R344 were proposed to be the potential key residues for catalysis via site-directed mutation. Detailed characterization of PMD and comprehensive comparisons could supply some different information about properties of PL 5 ALyases which might be helpful for its application in the directed production of AOS.

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“…The yield of trisaccharides was quantified as 758.60 mg/g based on the prepared standard curve (Figure 6C), and the proportion of trisaccharides in the AOS product, referred to as trisaccharide purity, was calculated to be 85.12%. There have been many studies on the production of AOS by alginate lyases, with degradation times typically spanning 6 to 48 h [23,30,40]. However, the yield of AOS has not been extensively quantified.…”

Section: Aos Preparation With the Produced Alyc7mentioning

confidence: 99%

“…In this study, we investigated the effect of the AOS produced by AlyC7 from sodium alginate on the root growth of different plants, including dicots, lettuce and tomato, and monocots, wheat and maize. As shown in Figure 7, compared to the control groups treated with distilled water, the root lengths of the seeds in the experimental groups treated with the AOS exhibited varying degrees There have been many studies on the production of AOS by alginate lyases, with degradation times typically spanning 6 to 48 h [23,30,40]. However, the yield of AOS has not been extensively quantified.…”

Section: Effects Of the Prepared Aos On The Root Growth Of Plant Seedsmentioning

confidence: 99%

High-Level Extracellular Production of a Trisaccharide-Producing Alginate Lyase AlyC7 in Escherichia coli and Its Agricultural Application

Wang,

Zhang,

Zhang

et al. 2024

Marine Drugs

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Alginate oligosaccharides (AOS), products of alginate degradation by endotype alginate lyases, possess favorable biological activities and have broad applications. Although many have been reported, alginate lyases with homogeneous AOS products and secretory production by an engineered host are scarce. Herein, the alginate lyase AlyC7 from Vibrio sp. C42 was characterized as a trisaccharide-producing lyase exhibiting high activity and broad substrate specificity. With PelB as the signal peptide and 500 mM glycine as the additive, the extracellular production of AlyC7 in Escherichia coli reached 1122.8 U/mL after 27 h cultivation in Luria-Bertani medium. The yield of trisaccharides from sodium alginate degradation by the produced AlyC7 reached 758.6 mg/g, with a purity of 85.1%. The prepared AOS at 20 μg/mL increased the root length of lettuce, tomato, wheat, and maize by 27.5%, 25.7%, 9.7%, and 11.1%, respectively. This study establishes a robust foundation for the industrial and agricultural applications of AlyC7.

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Construction and biochemical characterization of a novel hybrid alginate lyase with high activity by module recombination to prepare alginate oligosaccharides

Cited by 18 publications

References 30 publications

Production of Alginate Oligosaccharides (AOSs) Using Enhanced Physicochemical Properties of Immobilized Alginate Lyase for Industrial Application

Production of Alginate Oligosaccharides (AOSs) Using Enhanced Physicochemical Properties of Immobilized Alginate Lyase for Industrial Application

Characterization of a Novel Polysaccharide Lyase Family 5 Alginate Lyase with PolyM Substrate Specificity

High-Level Extracellular Production of a Trisaccharide-Producing Alginate Lyase AlyC7 in Escherichia coli and Its Agricultural Application

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