Study on expression and action mode of recombinant alginate lyases based on conserved domains reconstruction

Yang, Min; Li, Nannan; Yang, Suxiao; Yu, Yuan; Han, Zhong; Li, Li; Mou, Haijin

doi:10.1007/s00253-018-9502-7

Cited by 30 publications

(15 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As two examples, the specific activity of the recombinant alginate lyase Aly7B_Wf expressed in E. coli from Wenyingzhuangia fucanilytica was 23.24 U/mg [23] and the specific activity of AlyH1 from Vibrio furnissii H1 was 2.40 U/mg [11]. The specific activity of cAylM expressed in E. coli was 1386.27 U/mg [17], which was higher than that expressed in P. pastoris and it might be caused by the glycosylation modification of enzyme. The predicted glycosylation sites in cAlyM are shown in Figure S1.…”

Section: Purification Of Recombinant Calym and 102c300cmentioning

confidence: 94%

See 1 more Smart Citation

Expression and Characterization of an Alginate Lyase and Its Thermostable Mutant in Pichia pastoris

Yang

Liu

et al. 2020

Marine Drugs

Self Cite

View full text Add to dashboard Cite

Alginate is one of the most abundant polysaccharides in algae. Alginate lyase degrades alginate through a β-elimination mechanism to produce alginate oligosaccharides with special bioactivities. Improving enzyme activity and thermal stability can promote the application of alginate lyase in the industrial preparation of alginate oligosaccharides. In this study, the recombinant alginate lyase cAlyM and its thermostable mutant 102C300C were expressed and characterized in Pichia pastoris. The specific activities of cAlyM and 102C300C were 277.1 U/mg and 249.6 U/mg, respectively. Both enzymes showed maximal activity at 50 °C and pH 8.0 and polyG preference. The half-life values of 102C300C at 45 °C and 50 °C were 2.6 times and 11.7 times the values of cAlyM, respectively. The degradation products of 102C300C with a lower degree of polymerization contained more guluronate. The oligosaccharides with a polymerization degree of 2–4 were the final hydrolytic products. Therefore, 102C300C is potentially valuable in the production of alginate oligosaccharides with specific M/G ratio and molecular weights.

show abstract

Section: Purification Of Recombinant Calym and 102c300cmentioning

confidence: 94%

“…We previously reported that an alginate lyase cAlyM with high enzymatic activity from Microbulbifer sp. Q7 (CGMCC 14061) expressed in E. coli [17]. However, E. coli is unable to secrete recombinant protein into the medium, and the application of the recombinant protein produced by E. coli is limited due to the safety of residual endotoxin.…”

Section: Introductionmentioning

confidence: 99%

Expression and Characterization of an Alginate Lyase and Its Thermostable Mutant in Pichia pastoris

Yang

Liu

et al. 2020

Marine Drugs

Self Cite

View full text Add to dashboard Cite

show abstract

“…H1 [ 42 ] and AlyM from Microbulbifer sp. CGMCC 14061 [ 43 ] exhibit a completely different trait and display higher reactivity and thermal stability following loss of their CBM32 domain. However, the CBM32 domain of AlyQ from Persicobacter sp.…”

Section: Alginate Lyasesmentioning

confidence: 99%

Bacterial alginate metabolism: an important pathway for bioconversion of brown algae

Zhang

Xue

Zhang

et al. 2021

Biotechnol Biofuels

View full text Add to dashboard Cite

Brown macroalgae have attracted great attention as an alternative feedstock for biorefining. Although direct conversion of ethanol from alginates (major components of brown macroalgae cell walls) is not amenable for industrial production, significant progress has been made not only on enzymes involved in alginate degradation, but also on metabolic pathways for biorefining at the laboratory level. In this article, we summarise recent advances on four aspects: alginate, alginate lyases, different alginate-degrading systems, and application of alginate lyases and associated pathways. This knowledge will likely inspire sustainable solutions for further application of both alginate lyases and their associated pathways.

show abstract

“…The alginate lyase cAlyM belonging to PL7 from Microbulbifer sp. Q7 has been expressed in Escherichia coli , and its particular properties have also been depicted [22]. However, cAlyM cannot meet the demand for industrial preparation of alginate oligosaccharides, owing to its low thermal stability.…”

Section: Introductionmentioning

confidence: 99%

Rational Design of Alginate Lyase from Microbulbifer sp. Q7 to Improve Thermal Stability

Yang

Liu

et al. 2019

Marine Drugs

Self Cite

View full text Add to dashboard Cite

Alginate lyase degrades alginate by the β-elimination mechanism to produce oligosaccharides with special bioactivities. The low thermal stability of alginate lyase limits its industrial application. In this study, introducing the disulfide bonds while using the rational design methodology enhanced the thermal stability of alginate lyase cAlyM from Microbulbifer sp. Q7. Enzyme catalytic sites, secondary structure, spatial configuration, and molecular dynamic simulation were comprehensively analyzed. When compared with cAlyM, the mutants D102C-A300C and G103C-T113C showed an increase by 2.25 and 1.16 h, respectively, in half-life time at 45 °C, in addition to increases by 1.7 °C and 0.4 °C in the melting temperature, respectively. The enzyme-specific activity and kcat/Km values of D102C-A300C were 1.8- and 1.5-times higher than those of cAlyM, respectively. The rational design strategy that was used in this study provides a valuable method for improving the thermal stability of the alginate lyase.

show abstract

Study on expression and action mode of recombinant alginate lyases based on conserved domains reconstruction

Cited by 30 publications

References 38 publications

Expression and Characterization of an Alginate Lyase and Its Thermostable Mutant in Pichia pastoris

Expression and Characterization of an Alginate Lyase and Its Thermostable Mutant in Pichia pastoris

Bacterial alginate metabolism: an important pathway for bioconversion of brown algae

Rational Design of Alginate Lyase from Microbulbifer sp. Q7 to Improve Thermal Stability

Contact Info

Product

Resources

About