Enhanced a novel β-agarase production in recombinant Escherichia coli BL21 (DE3) through induction mode optimization and glycerol feeding strategy

CHAN Zhuhua; CHEN Xinglin; HOU Yanping; GAO Boliang; ZHAO Chungui; YANG Suping; ZENG Runying

doi:10.1007/s13131-018-1172-x

Article Contents

Article Navigation > Acta Oceanologica Sinica > 2018 > 37(2): 110-118

CHAN Zhuhua, CHEN Xinglin, HOU Yanping, GAO Boliang, ZHAO Chungui, YANG Suping, ZENG Runying. Enhanced a novel β-agarase production in recombinant Escherichia coli BL21 (DE3) through induction mode optimization and glycerol feeding strategy[J]. Acta Oceanologica Sinica, 2018, 37(2): 110-118. doi: 10.1007/s13131-018-1172-x

Citation:

CHAN Zhuhua, CHEN Xinglin, HOU Yanping, GAO Boliang, ZHAO Chungui, YANG Suping, ZENG Runying. Enhanced a novel β-agarase production in recombinant Escherichia coli BL21 (DE3) through induction mode optimization and glycerol feeding strategy[J]. Acta Oceanologica Sinica, 2018, 37(2): 110-118. doi: 10.1007/s13131-018-1172-x

Citation:

CHAN Zhuhua, CHEN Xinglin, HOU Yanping, GAO Boliang, ZHAO Chungui, YANG Suping, ZENG Runying. Enhanced a novel β-agarase production in recombinant Escherichia coli BL21 (DE3) through induction mode optimization and glycerol feeding strategy[J]. Acta Oceanologica Sinica, 2018, 37(2): 110-118. doi: 10.1007/s13131-018-1172-x

PDF( 396 KB)

Enhanced a novel β-agarase production in recombinant Escherichia coli BL21 (DE3) through induction mode optimization and glycerol feeding strategy

doi: 10.1007/s13131-018-1172-x

1.
Department of Bioengineering and Biotechnology, Huaqiao University, Xiamen 361021, China;State Key Laboratory Breeding Base of Marine Genetic Resource, Third Institute of Oceanography, State Oceanic Administration, Xiamen 361005, China
2.
State Key Laboratory Breeding Base of Marine Genetic Resource, Third Institute of Oceanography, State Oceanic Administration, Xiamen 361005, China
3.
State Key Laboratory Breeding Base of Marine Genetic Resource, Third Institute of Oceanography, State Oceanic Administration, Xiamen 361005, China;School of Life Sciences, Xiamen University, Xiamen 361102, China
4.
Department of Bioengineering and Biotechnology, Huaqiao University, Xiamen 361021, China

Received Date: 2016-12-28
Rev Recd Date: 2011-09-21

Abstract

Abstract

Agarases are hydrolytic enzymes that act on the hydrolysis of agar and have a broad range of applications in food, cosmetics and pharmaceutical industries. In this study, a glycerol feeding strategy based on induction mode optimization for high cell density and β-agarase production was established, which could effectively control acetate yield. First, exponential feeding strategy of glycerol with different overall specific growth rates (μ) was applied in the pre-induction phase. The results showed that the low μ (μ=0.2) was suggested to be the optimal for cell growth and β-agarase production. Second, the effects of induction temperature and the inducer concentration on cell growth and β-agarase production were investigated in the post-induction phase. When induced by isopropyl-β-d-thiogalactoside (IPTG), the strategy of 0.8 mmol/L IPTG induction at 20℃ was found to be optimal for β-agarase production. When cultivation was induced by continuous lactose feeding strategy of 1.0 g/(L·h), the β-agarase activity reached 112.5 U/mL, which represented the highest β-agarase production to date. Furthermore, the β-agarase was capable of degrading G. lemaneiformis powder directly to produce neoagarooligosaccharide, and the hydrolysates were neoagarotetraose (NA4) and neoagarohexaose (NA6). The overall research may be useful for the industrial production and application of β-agarase.
- β-agarase,
- Escherichia coli,
- process optimization,
- glycerol feeding strategy,
- neoagarooligosaccharide

FullText(HTML)

References(2)

References

Araki T, Hayakawa M, Lu Zhang, et al. 1998. Purification and characterization of agarases from a marine bacterium, Vibrio sp. PO-303. J Mar Biotechnol, 6(4): 260-265
Chan Zhuhua, Wang Runping, Liu Shenglong, et al. 2015. Draft genome sequence of an agar-degrading marine bacterium Flammeovirga pacifica WPAGA1. Mar Genom, 20: 23-24
Chen Xinglin, Hou Yanping, Jin Min, et al. 2016. Expression and characterization of a novel thermostable and pH-stable β-agarase from deep-sea bacterium Flammeovirga sp. OC4. J Agric Food Chem, 64(38): 7251-7258
Cheng Jing, Wu Dan, Chen Sheng, et al. 2011. High-level extracellular production of α-cyclodextrin glycosyltransferase with recombinant Escherichia coli BL21 (DE3). J Agric Food Chem, 59(8): 3797-3802
Chi W J, Park J S, Kang D K, et al. 2014. Production and characterization of a novel thermostable extracellular agarase from Pseudoalteromonas hodoensis newly isolated from the west sea of South Korea. Appl Biochem Biotechnol, 173(7): 1703-1716
Choi H J, Hong J B, Park J J, et al. 2011. Production of agarase from a novel Micrococcus sp. GNUM-08124 strain isolated from the East Sea of Korea. Biotechnol Bioprocess Eng, 16(1): 81-88
Dong Qi, Ruan Linwei, Shi Hong. 2016. A β-agarase with high pH stability from Flammeovirga sp. SJP92. Carbohyd Res, 432: 1-8
Dong Jinhua, Tamaru Y, Araki T. 2007. A unique β-agarase, AgaA, from a marine bacterium, Vibrio sp. strain PO-303. Appl Microbiol Biotechnol, 74(6): 1248-1255
Dvorak P, Chrast L, Nikel P I, et al. 2015. Exacerbation of substrate toxicity by IPTG in Escherichia coli BL21(DE3) carrying a synthetic metabolic pathway. Microb Cell Fact, 14: 201
Fang Shuying, Li Jianghua, Liu Long, et al. 2011. Overproduction of alkaline polygalacturonate lyase in recombinant Escherichia coli by a two-stage glycerol feeding approach. Bioresour Technol, 102(22): 10671-10678
Goyal D, Sahni G, Sahoo D K. 2009. Enhanced production of recombinant streptokinase in Escherichia coli using fed-batch culture. Bioresour Technol, 100(19): 4468-4474
Han Wenjun, Gu Jingyan, Yan Qiujie, et al. 2012. A polysaccharide-degrading marine bacterium Flammeovirga sp. MY04 and its extracellular agarase system. J Ocean Univ China, 11(3): 375-382
Hassairi I, Ben Amar R, Nonus M, et al. 2001. Production and separation of α-agarase from Altermonas agarlyticus strain GJ1B. Bioresour Technol, 79(1): 47-51
Hou Yanping, Chen Xinglin, Chan Zhuhua, et al. 2015. Expression and characterization of a thermostable and pH-stable β-agarase encoded by a new gene from Flammeovirga pacifica WPAGA1. Process Biochem, 50(7): 1068-1075
Jang M K, Lee D G, Kim N Y, et al. 2009. Purification and characterization of neoagarotetraose from hydrolyzed agar. J Microbiol Biotechnol, 19(10): 1197-1200

Jhamb K, Sahoo D K. 2012. Production of soluble recombinant proteins in Escherichia coli: effects of process conditions and chaperone co-expression on cell growth and production of xylanase. Bioresour Technol, 123: 135-143
Kilikian B V, Suárez I D, Liria C W, et al. 2000. Process strategies to improve heterologous protein production in Escherichia coli under lactose or IPTG induction. Process Biochem, 35(9): 1019-1025
Kobayashi R, Takisada M, Suzuki T, et al. 1997. Neoagarobiose as a novel moisturizer with whitening effect. Biosci Biotechnol Biochem, 61(1): 162-163
Lakshmikanth M, Manohar S, Patnakar J, et al. 2006a. Optimization of culture conditions for the production of extracellular agarases from newly isolated Pseudomonas aeruginosa AG LSL-11. World J Microbiol Biotechnol, 22(5): 531-537
Lakshmikanth M, Manohar S, Souche Y, et al. 2006b. Extracellular β-agarase LSL-1 producing neoagarobiose from a newly isolated agar-liquefying soil bacterium, Acinetobacter sp., AG LSL-1. World J Microbiol Biotechnol, 22(10): 1087-1094
Lee S B, Park J H, Yoon S C, et al. 2000. Sequence analysis of a β-agarase gene (pjaA) from Pseudomonas sp. isolated from Marine Environment. J Biosci Bioeng, 89(5): 485-488
Li Jiang, Sha Yujie. 2015. Expression and enzymatic characterization of a cold-adapted β-agarase from Antarctic bacterium Pseudoalteromonas sp. NJ21. Chin J Oceanol Limnol, 33(2): 319-327
Lin Bokun, Lu Guoyong, Zheng Yandan, et al. 2012. Gene cloning, expression and characterization of a neoagarotetraose-producing β-agarase from the marine bacterium Agarivorans sp. HZ105. World J Microbiol Biotechnol, 28(4): 1691-1697
Liu Yang, Yi Zhiwei, Cai Yaping, et al. 2015. Draft genome sequence of algal polysaccharides degradation bacterium, Flammeovirga sp. OC4. Mar Genom, 21: 21-22
Long Mengxian, Yu Ziniu, Xu Xun. 2010. A novel β-agarase with high pH stability from marine Agarivorans sp. LQ48. Mar Biotechnol, 12(1): 62-69
Malakar P, Venkatesh K V. 2012. Effect of substrate and IPTG concentrations on the burden to growth of Escherichia coli on glycerol due to the expression of Lac proteins. Appl Microbiol Biotechnol, 93(6): 2543-2549
Miller G L. 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem, 31(3): 426-428
Oh C, Nikapitiya C, Lee Y, et al. 2010. Cloning, purification and biochemical characterization of beta agarase from the marine bacterium Pseudoalteromonas sp. AG4. J Ind Microbiol Biotechnol, 37(5): 483-494
Ramalingam S, Gautam P, Mukherjee K J, et al. 2007. Effects of post-induction feed strategies on secretory production of recombinant streptokinase in Escherichia coli. Biochem Eng J, 33(1): 34-41
Roseline T L, Sachindra N M. 2016. Characterization of extracellular agarase production by Acinetobacter junii PS12B, isolated from marine sediments. Biocatal Agric Biotechnol, 6: 219-226
Seo Y B, Lu Yan, Chi W J, et al. 2014. Heterologous expression of a newly screened β-agarase from Alteromonas sp. GNUM1 in Escherichia coli and its application for agarose degradation. Process Biochem, 49(3): 430-436
Shokri A, Sandén A, Larsson G. 2003. Cell and process design for targeting of recombinant protein into the culture medium of Escherichia coli. Appl Microbiol Biotechnol, 60(6): 654-664
Sugano Y, Terada I, Arita M, et al. 1993. Purification and characterization of a new agarase from a marine bacterium, Vibrio sp. strain JT0107. Appl Environ Microbiol, 59(5): 1549-1554
Suzuki H, Sawai Y, Suzuki T, et al. 2003. Purification and characterization of an extracellular β-agarase from Bacillus sp. MK03. J Biosci Bioeng, 95(4): 328-334
Wang Huilin, Li Xiaoman, Ma Yanhe, et al. 2015. Process optimization of high-level extracellular production of alkaline pectate lyase in recombinant Escherichia coli BL21 (DE3). Biochem Eng J, 93: 38-46
Wang Jingxue, Mou Haijin, Jiang Xiaolu, et al. 2006. Characterization of a novel β-agarase from marine Alteromonas sp. SY37-12 and its degrading products. Appl Microbiol Biotechnol, 71(6): 833-839
Xie Wei, Lin Bokun, Zhou Zhengrong, et al. 2013. Characterization of a novel β-agarase from an agar-degrading bacterium Catenovulum sp. X3. Appl Microbiol Biotechnol, 97(11): 4907-4915
Xu Hui, Fu Yuanyuan, Yang Ning, et al. 2011. Flammeovirga pacifica sp. nov. , isolated from deep-sea sediment. Int J Syst Evol Microbiol, 62(4): 937-941
Xu Peng, Gu Qin, Wang Wenya, et al. 2013. Modular optimization of multi-gene pathways for fatty acids production in E. coli. Nat Commun, 4: 1409
Xu Zhinan, Liu Gang, Cen Peilin, et al. 2000. Factors influencing excretive production of human epidermal growth factor (hEGF) with recombinant Escherichia coli K12 system. Bioprocess Eng, 23(6): 669-674
Yang Jinglong, Chen L C, Shih Y Y, et al. 2011. Cloning and characterization of β-agarase AgaYT from Flammeovirga yaeyamensis strain YT. J Biosci Bioeng, 112(3): 225-232
Yang Meng, Mao Xiangzhao, Liu Nan, et al. 2014. Purification and characterization of two agarases from Agarivorans albus OAY02. Process Biochem, 49(5): 905-912
Yoshizawa Y, Ametani A, Tsunehiro J, et al. 1995. Macrophage stimulation activity of the polysaccharide fraction from a marine alga (Porphyra yezoensis): structure-function relationships and improved solubility. Biosci Biotechnol Biochem, 59(10): 1933-1937
Zhu Yanbing, Zhao Rui, Xiao Anfeng, et al. 2016. Characterization of an alkaline β-agarase from Stenotrophomonas sp. NTa and the enzymatic hydrolysates. Int J Biol Macromol, 86: 525-534

Relative Articles

Supplements(0)

Cited By

Proportional views