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

Lee, Yeong Rim; Jung, Subin; Chi, Won-Jae; Bae, Chang−Hwan; Jeong, Byeong Chul; Hong, Soon-Kwang; Lee, Chang-Ro

doi:10.4014/jmb.1710.10011

Cited by 18 publications

(10 citation statements)

References 33 publications

(69 reference statements)

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“…The agarases that shared similar properties to Ca Aga1 were characterized as AgaJ5 (GH86, from Gayadomonas joobiniege G7), AgaJ9 (GH39, from G. joobiniege G7), and Aga21 (from Pseudoalteromonas sp. NJ21), which had 40% activity at 10 °C [19], 80% activity at 5 °C [20], and 80% activity at 5 °C [21], respectively (Table 1). The cold-adapted feature enables Ca Aga1 to act in energy saving conditions (lowering the reaction temperature) and is of special interest for industrial applications.…”

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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Section: Discussionmentioning

confidence: 99%

Biochemical Characterization of a New β-Agarase from Cellulophaga algicola

Han

Zhang

Yang

2019

IJMS

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“…This agarase showed maximum activity at 45 °C and was stable below the same temperature (Xu et al 2018). The agarases, AgaJ5 and AgJ11 from G. joobiniege G7, were the only agarases found to be optimum in acidic pH 4.5 with the stability between pH of 4-5.5 (Lee et al 2018;Jung et al 2017a, b). Furthermore, the agarase, AgaP4383 from Flammeovirga pacifica WPAGA1, showed optimum activity at alkaline pH 9 and found to stable between pH 5 and 10 (Hou et al 2015).…”

Section: Physiochemical Properties Of the Recombinant β-Agarasesmentioning

confidence: 94%

“…In the presence of 200 mM of NaCl, AgaB1 exhibited its maximum activity (Liang et al 2014). Furthermore, the Na + and K + being present in the reaction mixture slightly activated the agarase activity of AgaJ5 (Lee et al 2018).…”

Section: Effect Of Metal Ions On β-Agarase Activitymentioning

confidence: 95%

“…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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“…However, only limited agarases with cold-adaptation, such as AgaJ5, AgaJ9, and Aga21, have been identified and characterized in recent years. In detail, AgaJ5 retained ~40% of the maximum enzymatic activity at 10 °C [ 13 ]. AgaJ9 maintained more than 80% of the maximum activity at 5 °C [ 8 ], and Aga21 retained as much as 85% of the maximum activity at 10 °C [ 14 ].…”

Section: Introductionmentioning

confidence: 99%

Expression and Characterization of a Novel Cold-Adapted and Stable β-Agarase Gene agaW1540 from the Deep-Sea Bacterium Shewanella sp. WPAGA9

Wang

Yan

et al. 2021

Marine Drugs

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The neoagaro-oligosaccharides, degraded from agarose by agarases, are important natural substances with many bioactivities. In this study, a novel agarase gene, agaW1540, from the genome of a deep-sea bacterium Shewanella sp. WPAGA9, was expressed, and the recombinant AgaW1540 (rAgaW1540) displayed the maximum activity under the optimal pH and temperature of 7.0 and 35 °C, respectively. rAgaW1540 retained 85.4% of its maximum activity at 0 °C and retained more than 92% of its maximum activity at the temperature range of 20–40 °C and the pH range of 4.0–9.0, respectively, indicating its extensive working temperature and pH values. The activity of rAgaW1540 was dramatically suppressed by Cu2+ and Zn2+, whereas Fe2+ displayed an intensification of enzymatic activity. The Km and Vmax of rAgaW1540 for agarose degradation were 15.7 mg/mL and 23.4 U/mg, respectively. rAgaW1540 retained 94.7%, 97.9%, and 42.4% of its maximum activity after incubation at 20 °C, 25 °C, and 30 °C for 60 min, respectively. Thin-layer chromatography and ion chromatography analyses verified that rAgaW1540 is an endo-acting β-agarase that degrades agarose into neoagarotetraose and neoagarohexaose as the main products. The wide variety of working conditions and stable activity at room temperatures make rAgaW1540an appropriate bio-tool for further industrial production of neoagaro-oligosaccharides.

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

Cited by 18 publications

References 33 publications

Biochemical Characterization of a New β-Agarase from Cellulophaga algicola

Biochemical Characterization of a New β-Agarase from Cellulophaga algicola

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

Expression and Characterization of a Novel Cold-Adapted and Stable β-Agarase Gene agaW1540 from the Deep-Sea Bacterium Shewanella sp. WPAGA9

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