Characterizing of a new α-agarase AgaE from Thalassomonas sp. LD5 and probing its catalytically essential residues

Xu, Jingnan; Cui, Zibo; Zhang, Wei-Bin; Lu, Jingxuan; Lu, Xin; Yu, Wengong

doi:10.1016/j.ijbiomac.2021.11.194

Cited by 6 publications

(6 citation statements)

References 29 publications

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“…Compared with the reported α‐agarases CaLJ96 and OUC‐GaJJ96, which almost inactivated after incubating at 35°C and 37°C for 1 h, the recombinant Cm‐AGA showed better thermal stability (Jin et al, 2020; Liu et al, 2019). Meanwhile, Cm‐AGA exhibited identical optimal reaction temperature but weaker thermal stability compared with AgaD and AgaE (J. Xu et al, 2022; W. Zhang, Xu, et al, 2018), probably caused by the relatively large molecular weight of Cm‐AGA and the wide distribution of highly flexible regions (Ding et al, 2021).…”

Section: Resultsmentioning

confidence: 99%

“…Meanwhile, Cm-AGA exhibited identical optimal reaction temperature but weaker thermal stability compared with AgaD and AgaE (J. Xu et al, 2022;W. Zhang, Xu, et al, 2018), probably caused by the relatively large molecular weight of Cm-AGA and the wide distribution of highly flexible regions (Ding et al, 2021).…”

Section: Characterization Of Recombinant Cm-agamentioning

confidence: 99%

“…The α-agarase CaLJ96 could produce A2, A4, and A6 with a proportion of 1:6.91:3.14 (Liu et al, 2019). Both AgaA and AgaE degraded the agarose to produce A4 and A6, whereas the dominant product of AgaA was A4 and that of AgaE was A6 (Potin et al, 1993;J. Xu et al, 2022).…”

Section: Degradation Product Analysismentioning

confidence: 99%

“…However, only seven α‐agarases have been characterized to date, including AgaA from Alteromonas agarilytica GJ1B (Hassairi et al, 2001), AgaA33 from Thalassotalea agarivorans JAMB A33 (Ohta et al, 2005), AgaWS5 from Catenovulum sediminis WS1‐A (Lee et al, 2019), OUC‐GaJJ96 from Gilvimarinus agarilyticus (Jin et al, 2020), CaLJ96 from Catenovulum agarivorans (Liu et al, 2019), and AgaD and AgaE from Thalassomonas sp. LD5 (Wang et al, 2020; J. Xu et al, 2022). All the reported α‐agarases are classified into the GH96 family based on the amino acid sequence similarity.…”

Section: Introductionmentioning

confidence: 99%

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Characterization and efficient production of an α‐agarase from marine bacterium Catenovulum maritimum STB14

You

Xie

et al. 2022

Food Bioengineering

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Agarases are enzymes that degrade agar and agarose to produce agar oligosaccharides with multiple functional activities. Compared with β‐agarases, the natural source of α‐agarases is limited, which severely restricts the industrial application of α‐agarases. Here, we cloned and heterologously expressed an α‐agarase belonging to glycoside hydrolase family 96 named Cm‐AGA from the marine bacterium Catenovulum maritimum STB14. The production conditions of recombinant Cm‐AGA were optimized as: taking Terrific Broth (TB) (pH 6.5) with 5 g/L of fructose and 24 g/L of yeast extract H07014 as the fermentation medium, after culturing at 37°C for 2 h, isopropyl‐β‐ d‐thiogalactoside was added with a final concentration of 0.01 mM to induce for 44 h. The obtained enzyme activity was 13.81 U/ml and was about 6.6 times the initial activity. The specific activity of recombinant Cm‐AGA was 206.1 U/mg, the optimum temperature and pH were 35°C and 8.0, respectively, and the enzyme activity could be activated by Mn2+ and Ca2+. The hydrolysis product results showed that Cm‐AGA is the first reported α‐agarase with agarobiose (A2) and agarotetraose (A4) as the dominant products, suggesting the great potential of Cm‐AGA in the efficient production of agaro‐oligosaccharides with a low degree of polymerization.

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Section: Resultsmentioning

confidence: 99%

Section: Characterization Of Recombinant Cm-agamentioning

confidence: 99%

Section: Degradation Product Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Characterization and efficient production of an α‐agarase from marine bacterium Catenovulum maritimum STB14

You

Xie

et al. 2022

Food Bioengineering

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“…The strain Thalassomonas sp. LD5 capable of degrading agar [ 40 , 41 ], alginate [ 42 , 43 ], and ulvan, was screened from Qingdao offshore. Restriction enzymes pET-28a (+) vector plasmid (from Takara Co., Ltd., Dalian, China), T4 ligase, restriction endonuclease Nde I and Xho I, and DNA polymerase (from Vazyme Biotech Co., Ltd., Nanjing, China) were used for Vector construction.…”

Section: Methodsmentioning

confidence: 99%

Biochemical Properties of a New Polysaccharide Lyase Family 25 Ulvan Lyase TsUly25B from Marine Bacterium Thalassomonas sp. LD5

Wang

Han

et al. 2022

Marine Drugs

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Marine macroalgae, contributing much to the bioeconomy, have inspired tremendous attention as sustainable raw materials. Ulvan, as one of the main structural components of green algae cell walls, can be degraded by ulvan lyase through the β-elimination mechanism to obtain oligosaccharides exhibiting several good physiological activities. Only a few ulvan lyases have been characterized until now. This thesis explores the properties of a new polysaccharide lyase family 25 ulvan lyase TsUly25B from the marine bacterium Thalassomonas sp. LD5. Its protein molecular weight was 54.54 KDa, and it was most active under the conditions of 60 °C and pH 9.0. The Km and kcat values were 1.01 ± 0.05 mg/mL and 10.52 ± 0.28 s−1, respectively. TsUly25B was salt-tolerant and NaCl can significantly improve its thermal stability. Over 80% of activity can be preserved after being incubated at 30 °C for two days when the concentration of NaCl in the solution is above 1 M, while 60% can be preserved after incubation at 40 °C for 10 h with 2 M NaCl. TsUly25B adopted an endolytic manner to degrade ulvan polysaccharides, and the main end-products were unsaturated ulvan disaccharides and tetrasaccharides. In conclusion, our research enriches the ulvan lyase library and advances the utilization of ulvan lyases in further fundamental research as well as ulvan oligosaccharides production.

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The α-linkage in funoran and agarose could be hydrolyzed by a GH96 family enzyme: Discovery of the α-funoranase

Zhang,

Zheng,

Liu

et al. 2024

Carbohydrate Polymers

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Characterizing of a new α-agarase AgaE from Thalassomonas sp. LD5 and probing its catalytically essential residues

Cited by 6 publications

References 29 publications

Characterization and efficient production of an α‐agarase from marine bacterium Catenovulum maritimum STB14

Characterization and efficient production of an α‐agarase from marine bacterium Catenovulum maritimum STB14

Biochemical Properties of a New Polysaccharide Lyase Family 25 Ulvan Lyase TsUly25B from Marine Bacterium Thalassomonas sp. LD5

The α-linkage in funoran and agarose could be hydrolyzed by a GH96 family enzyme: Discovery of the α-funoranase

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