Characterization and overexpression of a glycosyl hydrolase family 16 beta-agarase YM01-1 from marine bacterium Catenovulum agarivorans YM01 T

An, Ke; Shi, Xiaochong; Cui, Fangyuan; Cheng, Jingguang; Liu, Na; Zhao, Xia; Zhang, Xiaohua

doi:10.1016/j.pep.2017.10.002

Cited by 23 publications

(13 citation statements)

References 49 publications

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“…Actually, there were also some other reported agarases, of which the activities were not affected by urea. For example, agarase ID2563 could resist to 5 mM urea [26] and agarases YM01-1 and YM01-3 were resistant to 10 mM urea [30,31].…”

Section: Discussionmentioning

confidence: 99%

A new neoagarobiose-producing agarase from Vibrio sp. LA1

Lin¹,

Zheng²,

Huang³

et al. 2021

ScienceAsia

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An agarolytic bacterium was isolated from the sea coast of Shantou in China and identified as Vibrio sp. LA1. The agarase gene agaA was cloned from Vibrio sp. LA1 by using degenerate oligonucleotide-primed PCR and genome walking technique. Gene agaA consists of a 2913 bp open reading frame encoding 970 amino acids; and the predicted molecular mass and isoelectric point were 108 kDa and 4.46, respectively. Based on the amino acid sequence similarity, the encoded protein (AgaA) of gene agaA should be an agarase of glycoside hydrolase family GH50 with a catalytic domain of glycoside hydrolase family GH42. Soluble expression of AgaA in Escherichia coli was obtained and investigated. The optimal temperature and pH for the activity of the purified recombinant agarase were 35°C and pH 6, respectively. The reducing reagent β-mercaptoethanol could increase the activity of agarase AgaA by more than 80%. AgaA showed exo-lytic activity on agarose degradation. It decomposed agarose to yield neoagarobiose as the sole product, which was different from the agarase Aga41A from Vibrio sp. CN41 that showed 95% identities of amino acid sequence to agarase AgaA. With single end product, purification procedure is easier than that with multi-products. Therefore, agarase AgaA could be a useful tool for producing the bioactive neoagarobiose.

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Section: Discussionmentioning

confidence: 99%

A new neoagarobiose-producing agarase from Vibrio sp. LA1

Lin¹,

Zheng²,

Huang³

et al. 2021

ScienceAsia

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“…The NA2-producing agarases included N3-1 from Microbulbifer sp. BN3 [32], YM01-1 from marine bacterium C. agarivorans YM01 [33], AgaH71 from Pseudoalteromonas hodoensis [34], AgaG1 from Alteromonas sp. GNUM-1 [35], and Zg AgaA and Zg AgaB from Z. galactanivorans Dsij [36,37] (Table 1).…”

Section: Discussionmentioning

confidence: 99%

Biochemical Characterization of a New β-Agarase from Cellulophaga algicola

Han

Zhang

Yang

2019

IJMS

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Cellulophaga algicola DSM 14237, isolated from the Eastern Antarctic coastal zone, was found to be able to hydrolyze several types of polysaccharide materials. In this study, a predicted β-agarase (CaAga1) from C. algicola was heterologously expressed in Escherichia coli. The purified recombinant CaAga1 showed specific activities of 29.39, 20.20, 14.12, and 8.99 U/mg toward agarose, pure agar, and crude agars from Gracilaria lemaneiformis and Porphyra haitanensis, respectively. CaAga1 exhibited an optimal temperature and pH of 40 oC and 7, respectively. CaAga1 was stable over a wide pH range from 4 to 11. The recombinant enzyme showed an unusual thermostability, that is, it was stable at temperature below or equal to 40oC and around 70 oC, but was thermolabile at about 50 oC. With the agarose as the substrate, the Km and Vmax values for CaAga1 were 1.19 mg/mL and 36.21 U/mg, respectively. The reducing reagent (dithiothreitol) enhanced the activity of CaAga1 by more than one fold. In addition, CaAga1 was salt-tolerant given that it retained approximately 70% of the maximum activity in the presence of 2 M NaCl. The thin layer chromatography results indicated that CaAga1 is an endo-type β-agarase and efficiently hydrolyzed agarose into neoagarotetraose (NA4) and neoagarohexaose (NA6). A structural model of CaAga1 in complex with neoagarooctaose (NA8) was built by homology modeling and explained the hydrolysis pattern of CaAga1.

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“…The protein denaturant urea contradictorily did not affect the agarases ID2563, YM01-1, and YM01-3 (Rajagopalan et al 1961). In point of fact, the agarase ID2563 was resistant to urea and the agarase activity of YM01-1 and YM01-3 was not affected by urea (Su et al 2017;An et al 2018;Cui et al 2014). However, the presence of urea negatively influenced the agarase AgaM1 (Di et al 2018).…”

Section: Effect Of Metal Ions On β-Agarase Activitymentioning

confidence: 93%

“…However, the presence of urea negatively influenced the agarase AgaM1 (Di et al 2018). The monovalent cation Ni + hardly had any effect on the agarases YM01-1 and AgaJ9 (An et al 2018;Jung et al 2017).…”

Section: Effect Of Metal Ions On β-Agarase Activitymentioning

confidence: 98%

“…The recombinant β-agarases belong to the glycoside hydrolase (GH) family such as GH16, GH39, GH50, GH86, and GH118. Predominantly, the recombinant β-agarases were from marine microbes and their genera include Agarivorans (Liu et al 2014a, b;Lee et al 2012;Lin et al 2012), Alteromonas (Seo et al 2014;Chi et al 2014), Aquimarina (Lin et al 2017), Catenovulum (An et al 2018;Cui et al 2014;Xie et al 2013), Cellulophaga (Ramos et al 2018), Flammeovirga (Dong et al 2016;Hou et al 2015;Yang et al 2011;Chen et al 2016;Di et al 2018), Gayadomonas (Lee et al 2018;Jung et al 2017a, b), Microbulbifer (Su et al 2017), Micrococcaceae (Xu et al 2018), Pseudoalteromonas (Chi et al 2015a, b;Oh et al 2010a, b), Pseudomonas (Hsu et al 2015), Saccharophagus (Kim et al 2010(Kim et al , 2017(Kim et al , 2018Lee et al 2013), Simiduia (Tawara et al 2015), Thalassomonas (Liang et al 2014), and Vibrio (Liao et al 2011) whereas few were from soil bacteria Streptomyces (Temuujin et al 2011(Temuujin et al , 2012, Cohnella (Li et al 2015) and one exclusive source was the activated sludges of sewage plant from which Cellvibrio (Osamu et al 2012) had been identified with agarolytic property.…”

Section: Introductionmentioning

confidence: 99%

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Recombinant β-agarases: insights into molecular, biochemical, and physiochemical characteristics

Veerakumar

Manian

2018

3 Biotech

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Agarases (agarose 4-glycanohydrolase; EC 3.2.1.81) are class of enzymes that belong to glycoside hydrolase (GH) family capable of hydrolyzing agar. Their classification depends on hydrolysis pattern and product formation. Among all the agarases, β-agarases and the oligosaccharides formed by its action have fascinated quite a lot of industries. Ample of β-agarase genes have been endowed from marine sources such as algae, sea water, and marine sediments, and the expression of these genes into suitable host gives rise to recombinant β-agarases. These recombinant β-agarases have wide range of industrial applications due to its improved catalytic efficiency and stability in tough environments with ease of production on large scale. In this review, we have perused different types of recombinant β-agarases in consort with their molecular, physiochemical, and kinetic properties in detail and the significant features of those agarases are spotlighted. From the literature reviewed after 2010, we have found that the recombinant β-agarases belonged to the families GH16, GH39, GH50, GH86, and GH118. Among that, GH39, GH50, and GH86 belonged to clan GH-A, while the GH16 family belonged to clan GH-B. It was observed that GH16 is the largest polyspecific glycoside hydrolase family with ample number of β-agarases and the families GH50 and GH118 were found to be monospecific with only β-agarase activity. And, out of 84 non-catalytic carbohydrate-binding modules (CBMs), only CBM6 and CBM13 were professed in β-agarases. We witnessed a larger heterogeneity in molecular, physiochemical, and catalytic characteristics of the recombinant β-agarases including molecular mass: 32-132 kDa, optimum pH: 4.5-9, optimum temperature 16-60 °C, K M : 0.68-59.8 mg/ml, and V max : 0.781-11,400 U/ mg. Owing to this extensive range of heterogeneity, they have lion's share in the multibillion dollar enzyme market. This review provides a holistic insight to a few aspects of recombinant β-agarases which can be referred by the upcoming explorers to this area.

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Characterization and overexpression of a glycosyl hydrolase family 16 beta-agarase YM01-1 from marine bacterium Catenovulum agarivorans YM01 T

Cited by 23 publications

References 49 publications

A new neoagarobiose-producing agarase from Vibrio sp. LA1

A new neoagarobiose-producing agarase from Vibrio sp. LA1

Biochemical Characterization of a New β-Agarase from Cellulophaga algicola

Recombinant β-agarases: insights into molecular, biochemical, and physiochemical characteristics

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