Saccharification of Brown Macroalgae Using an Arsenal of Recombinant Alginate Lyases: Potential Application in the Biorefinery Process

Gimpel, Javier; Ravanal, María Cristina; Salazar, Osvaldo; Lienqueo, María Elena

doi:10.4014/jmb.1805.05056

Cited by 8 publications

(6 citation statements)

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“…Nearly half of known alginate lyase sequences have been classified into the PL7 family; however, exo-type PL7 alginate lyases have rarely been reported [18,39,44,45]. In this paper, the exo-type PL7 alginate lyase VxAly7D from V. xiamenensis QY104 was cloned and characterized.…”

Section: Discussionmentioning

confidence: 99%

“…Some of these enzymes even show apparent substrate preferences [32,[35][36][37], which limits the choice of possible substrates and affects the process of industrialization. To efficiently degrade alginate to produce unsaturated monosaccharides, it has recently been proposed to exploit the synergistic effects of endolytic and exolytic alginate lyases [4,6,38,39]. Nevertheless, the low activity of exo-type alginate lyases is still the rate-limiting step for obtaining unsaturated monosaccharides.…”

Section: Introductionmentioning

confidence: 99%

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Biochemical characteristics and molecular mechanism of an exo-type alginate lyase VxAly7D and its use for the preparation of unsaturated monosaccharides

Tang

Wang

Gao

et al. 2020

Biotechnol Biofuels

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Background As the most abundant polysaccharide in brown algae, alginate has become a promising economical material for bioethanol production. Recently, exo-type alginate lyases have received extensive attention because the unsaturated monosaccharides produced by their degradation of alginate can be easily converted into 4-deoxy-l-erythro-5-hexoseulose uronate (DEH), a promising material for bioethanol production and biorefinery systems. Results In this study, we cloned and characterized an exo-type polysaccharide lyase family 7 (PL7) alginate lyase VxAly7D from the marine bacterium Vibrio xiamenensis QY104. Recombinant VxAly7D was most active at 30 °C and exhibited 21%, 46% and 90% of its highest activity at 0, 10 and 20 °C, respectively. Compared with other exo-type alginate lyases, recombinant VxAly7D was shown to be a bifunctional alginate lyase with higher specific activity towards sodium alginate, polyG and polyM (462.4 ± 0.64, 357.37 ± 0.53 and 441.94 ± 2.46 U/mg, respectively). A total of 13 μg recombinant VxAly7D could convert 3 mg sodium alginate to unsaturated monosaccharides in 1 min with a yield of 37.6%, and the yield reached 95% in 1 h. In addition, the three-dimensional structure of VxAly7D was modelled using the crystal structure of AlyA5 from Zobellia galactanivorans DsijT as the template. The action mode and the end products of the W295A mutant revealed that Trp295 is a key amino acid residue responsible for the exolytic action mode of VxAly7D. Conclusion Overall, our results show that VxAly7D is a PL7 exo-type alginate lyase with high activity and a high conversion rate at low/moderate temperatures, which provides a useful enzymatic tool for the development of biofuel production from brown algae and enriches the understanding of the structure and functional relationships of polysaccharide lyases.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Biochemical characteristics and molecular mechanism of an exo-type alginate lyase VxAly7D and its use for the preparation of unsaturated monosaccharides

Tang

Wang

Gao

et al. 2020

Biotechnol Biofuels

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“…By contrast, a detailed study on the synergistic mechanism of alginate lyases has rarely been reported. Gimpel et al [31] used 21 different endo-/exo-lyase combinations to investigate the synergistic effect of different pairs, including 7 endo-lyases and 3 exo-lyases. However, they did not investigate the synergistic mechanism of these alginate lyases, while focused on only optimizing the conditions of the enzymatic reactions.…”

Section: Discussionmentioning

confidence: 99%

“…Endolytic and exolytic alginate lyases often co-exist in the same genome and are thus thought to work synergistically to cause the fast and complete digestion and utilization of alginate by bacteria. However, the synergistic mechanisms of endolytic and exolytic alginate lyases from the same bacterium strain remain to be investigated in detail, although a few studies have attempted to exploit the synergistic effect of endolytic and exolytic alginate lyases to maximize the alginate saccharification rate by optimizing the conditions of the enzymatic reaction [30, 31]. The biochemical characteristics and crystal structures of an endolytic alginate lyase, AlyA1, and an exolytic alginate lyase, AlyA5, from the marine flavobacterium Zobellia galactanivorans were studied in detail, but their synergistic effect was not investigated [32].…”

Section: Introductionmentioning

confidence: 99%

Biochemical characteristics and synergistic effect of two novel alginate lyases from Photobacterium sp. FC615

Zhang

Wang

et al. 2019

Biotechnol Biofuels

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BackgroundMacroalgae and microalgae, as feedstocks for third-generation biofuel, possess competitive strengths in terms of cost, technology and economics. The most important compound in brown macroalgae is alginate, and the synergistic effect of endolytic and exolytic alginate lyases plays a crucial role in the saccharification process of transforming alginate into biofuel. However, there are few studies on the synergistic effect of endolytic and exolytic alginate lyases, especially those from the same bacterial strain.ResultsIn this study, the endolytic alginate lyase AlyPB1 and exolytic alginate lyase AlyPB2 were identified from the marine bacterium Photobacterium sp. FC615. These two enzymes showed quite different and novel enzymatic properties whereas behaved a strong synergistic effect on the saccharification of alginate. Compared to that when AlyPB2 was used alone, the conversion rate of alginate polysaccharides to unsaturated monosaccharides when AlyPB1 and AlyPB2 acted on alginate together was dramatically increased approximately sevenfold. Furthermore, we found that AlyPB1 and AlyPB2 acted the synergistic effect basing on the complementarity of their substrate degradation patterns, particularly due to their M-/G-preference and substrate-size dependence. In addition, a novel method for sequencing alginate oligosaccharides was developed for the first time by combining the 1H NMR spectroscopy and the enzymatic digestion with the exo-lyase AlyPB2, and this method is much simpler than traditional methods based on one- and two-dimensional NMR spectroscopy. Using this strategy, the sequences of the final tetrasaccharide and pentasaccharide product fractions produced by AlyPB1 were easily determined: the tetrasaccharide fractions contained two structures, ΔGMM and ΔMMM, at a molar ratio of 1:3.2, and the pentasaccharide fractions contained four structures, ΔMMMM, ΔMGMM, ΔGMMM, and ΔGGMM, at a molar ratio of ~ 1:1.5:3.5:5.25.ConclusionsThe identification of these two novel alginate lyases provides not only excellent candidate tool-type enzymes for oligosaccharide preparation but also a good model for studying the synergistic digestion and saccharification of alginate in biofuel production. The novel method for oligosaccharide sequencing described in this study will offer a very useful approach for structural and functional studies on alginate.

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“…Fucoidan, as well as some polyphenols, from brown algae can be used for the production of pharmaceuticals, nutraceuticals and cosmetics, owing to their high bioactivity potential [ 16 , 17 , 18 ]. Alginate, a macroalgal hydrocolloid, is mainly used in food, pharmaceutical and medical applications, as well as in textile and paper production [ 19 , 20 , 21 ]. Furthermore, its gelling and thickening properties have been increasingly exploited to generate innovative biofilms and hydrogels [ 22 , 23 ].…”

Section: Introductionmentioning

confidence: 99%

Characterization and Bioactivity of Polysaccharides Separated through a (Sequential) Biorefinery Process from Fucus spiralis Brown Macroalgae

et al. 2022

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Marine macroalgae biomass is a valuable renewable resource that can be used for the development of bioeconomy through the valorisation of valuable compounds. The aim of the current study is separate macroalgal polysaccharides with bioactive properties from brown macroalgae Fucus spiralis based on a designed biocascading biorefinery approach. Thus, we applied an integrated processing method for the separation of fucoidan and alginate, in addition to characterization through IR spectroscopy and 1H NMR. The bioactivity potential (antioxidant activity using superoxide anion and DPPH radical scavenging analysis) of the two polysaccharides was evaluated, together with DNA binding studies performed though voltametric techniques and electronic spectroscopy titration. In terms of results, functional groups S=O (1226 cm−1), N=S=O (1136 cm−1) and C-O-SO3 (1024 cm−1), which are characteristic of fucoidan, were identified in the first polysaccharidic extract, whereas guluronic units (G) (1017 cm−1) and mannuronic units (M) (872 and 812 cm−1) confirmed the separation of alginate. The DNA binding studies of the isolated polysaccharides revealed an electrostatic and an intercalation interaction of DNA with fucoidan and alginate, respectively. Both antioxidant activity assays revealed improved antioxidant activity for both fucoidan and alginate compared to the standard α-tocopherol.

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Saccharification of Brown Macroalgae Using an Arsenal of Recombinant Alginate Lyases: Potential Application in the Biorefinery Process

Cited by 8 publications

References 65 publications

Biochemical characteristics and molecular mechanism of an exo-type alginate lyase VxAly7D and its use for the preparation of unsaturated monosaccharides

Biochemical characteristics and molecular mechanism of an exo-type alginate lyase VxAly7D and its use for the preparation of unsaturated monosaccharides

Biochemical characteristics and synergistic effect of two novel alginate lyases from Photobacterium sp. FC615

Characterization and Bioactivity of Polysaccharides Separated through a (Sequential) Biorefinery Process from Fucus spiralis Brown Macroalgae

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