Identification and biochemical characterization of a novel endo-type β-agarase AgaW from Cohnella sp. strain LGH

Li, Gen; Sun, Mingming; Jun, Wu; Ye, Mao; Ge, Xincheng; Wei, Wei; Li, Huixin; Hu, Feng

doi:10.1007/s00253-015-6869-6

Cited by 34 publications

(28 citation statements)

References 39 publications

(47 reference statements)

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“…AKV62624) from the soil bacterium Cohnella sp. LGH (Li et al 2015 ), which belongs to the GH50 family. The degree of identity and similarity between AgaB-4 and AgaW was 93.6 and 97.5%, respectively.…”

Section: Resultsmentioning

confidence: 99%

Molecular cloning, expression, and functional characterization of the β-agarase AgaB-4 from Paenibacillus agarexedens

et al. 2018

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In this study, a β-agarase gene, agaB-4, was isolated for the first time from the agar-degrading bacterium Paenibacillus agarexedens BCRC 17346 by using next-generation sequencing. agaB-4 consists of 2652 bp and encodes an 883-amino acid protein with an 18-amino acid signal peptide. agaB-4 without the signal peptide DNA was cloned and expressed in Escherichia coli BL21(DE3). His-tagged recombinant AgaB-4 (rAgaB-4) was purified from the soluble fraction of E. coli cell lysate through immobilized metal ion affinity chromatography. The optimal temperature and pH of rAgaB-4 were 55 °C and 6.0, respectively. The results of a substrate specificity test showed that rAgaB-4 could degrade agar, high-melting point agarose, and low-melting point agarose. The Vmax and Km of rAgaB-4 for low-melting point agarose were 183.45 U/mg and 3.60 mg/mL versus 874.61 U/mg and 9.29 mg/mL for high-melting point agarose, respectively. The main products of agar and agarose hydrolysis by rAgaB-4 were confirmed to be neoagarotetraose. Purified rAgaB-4 can be used in the recovery of DNA from agarose gels and has potential application in agar degradation for the production of neoagarotetraose.Electronic supplementary materialThe online version of this article (10.1186/s13568-018-0581-8) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Molecular cloning, expression, and functional characterization of the β-agarase AgaB-4 from Paenibacillus agarexedens

et al. 2018

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“…Over and above, some of the metal ions had inhibitory or negative effects on the enzyme activity of the agarases. The partial inhibition of agarase activity by Ba 2+ ions was detected in AgaW (Li et al 2015). The Sr + and Ba 2+ ions inhibited the activity of Aga4436 (Chen et al 2016).…”

Section: Effect Of Metal Ions On β-Agarase Activitymentioning

confidence: 94%

“…A handful of metal ions positively influenced the activity of a few β-agarases. The redox reagent dithiothreitol (DTT) had stimulatory effects on the agarases AgaB, AgaW, AgaXa, AgaM1, and HZ2 (Dong et al 2016;Li et al 2015;Xie et al 2013;Di et al 2018;Lin et al 2012). At higher concentrations of 9 mM and 10 mM, DTT increased the activity of HZ2 and AgaXa respectively (Lin et al 2012;Xie et al 2013).…”

Section: Effect Of Metal Ions On β-Agarase Activitymentioning

confidence: 99%

“…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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“…Alteromonas [11], Pseudomonas [12], Gayadomonas [3], Vibrio [13,14], Pseudoalteromonas [15], Zobellia [16], Catenovulum [17], Agarivorans [18], Cohnella [19], Bacillus [20], Thalassomonas [21], Streptomyces [22,23], Stenotrophomonas [24], Simiduia [25], and Flammeovirga [26]. They have been classified into a number of glycoside hydrolase (GH) families, including GH16 [27], GH39 [3], GH50 [26], GH86 [28], and GH118 [18].…”

Section: Introductionmentioning

confidence: 99%

Biochemical Characterization of a Novel GH86 ��-Agarase Producing Neoagarohexaose from Gayadomonas joobiniege G7

Lee

Jung

Chi

et al. 2018

Journal of Microbiology and Biotechnology

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A novel β-agarase, AgaJ5, was identified from an agar-degrading marine bacterium, G7. It belongs to the glycoside hydrolase family 86 and is composed of 805 amino acids with a 30-amino-acid signal peptide. Zymogram analysis showed that purified AgaJ5 has agarase activity. The optimum temperature and pH for AgaJ5 activity were determined to be 30°C and 4.5, respectively. AgaJ5 was an acidic β-agarase that had strong activity at a narrow pH range of 4.5-5.5, and was a cold-adapted enzyme, retaining 40% of enzymatic activity at 10°C. AgaJ5 required monovalent ions such as Na and K for its maximum activity, but its activity was severely inhibited by several metal ions. The and of AgaJ5 for agarose were 8.9 mg/ml and 188.6 U/mg, respectively. Notably, thin-layer chromatography, mass spectrometry, and agarose-liquefication analyses revealed that AgaJ5 was an endo-type β-agarase producing neoagarohexaose as the final main product of agarose hydrolysis. Therefore, these results suggest that AgaJ5 from G7 is a novel endo-type neoagarohexaose-producing β-agarase having specific biochemical features that may be useful for industrial applications.

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Identification and biochemical characterization of a novel endo-type β-agarase AgaW from Cohnella sp. strain LGH

Cited by 34 publications

References 39 publications

Molecular cloning, expression, and functional characterization of the β-agarase AgaB-4 from Paenibacillus agarexedens

Molecular cloning, expression, and functional characterization of the β-agarase AgaB-4 from Paenibacillus agarexedens

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

Biochemical Characterization of a Novel GH86 ��-Agarase Producing Neoagarohexaose from Gayadomonas joobiniege G7

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