Applied Microbiology and Biotechnology

, Volume 102, Issue 12, pp 5165–5172 | Cite as

A novel agaro-oligosaccharide-lytic β-galactosidase from Agarivorans gilvus WH0801

  • Xiaoqing Yang
  • Zhen Liu
  • Chengcheng Jiang
  • Jianan Sun
  • Changhu Xue
  • Xiangzhao Mao
Biotechnologically relevant enzymes and proteins


β-Galactosidases have a wide application in the food and pharmaceutical industries. Recently, β-galactosidase was also found to participate in agar degradation. In this study, the second reported agarolytic β-galactosidase was found in the marine bacterium Agarivorans gilvus WH0801 and characterized. The β-galactosidase named AgWH2A (83 kDa) exhibited good activities under optimal hydrolysis conditions of pH 8.0 and 40 °C. AgWH2A could cleave the first D-galactose of agarooligosaccharides from its nonreducing end to produce neoagarooligosaccharides, but could not act on the neoagarooligosaccharides. AgWH2A has great potential in the comprehensive utilization of marine red algae.


β-Galactosidase GH2 family Agarivorans gilvus D-Galactose Agarooligosaccharides 


Funding information

This work was supported by the National Natural Science Foundation of China (31471607) and the Applied Basic Research Program of Qingdao (16-5-1-17-jch).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.


  1. Ariki C, Arai K (1957) Studies on the chemical constitution of agar-agar. XX. Isolation of a tetrasaccharide by enzymatic hydrolysis of agar-agar. Bull Chem Soc Jpn 30:287–293. CrossRefGoogle Scholar
  2. Bertani G (1951) Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. J Bacteriol 62:293–300PubMedPubMedCentralGoogle Scholar
  3. Białkowska AM, Cieśliński H, Nowakowska KM, Kur J, Turkiewicz M (2009) A new β-galactosidase with a low temperature optimum isolated from the Antarctic Arthrobacter sp. 20B: gene cloning, purification and characterization. Arch Microbiol 191:825–835. CrossRefPubMedGoogle Scholar
  4. Blanchard JE, Gal L, He S, Foisy J, Warren RAJ, Withers SG (2001) The identification of the catalytic nucleophiles of two β-galactosidases from glycoside hydrolase family 35. Carbohydr Res 333:7–17. CrossRefPubMedGoogle Scholar
  5. Campuzano S, Serra B, Llull D, García JL, García P (2009) Cloning, expression, and characterization of a peculiar choline-binding β-galactosidase from Streptococcus mitis. Appl Environ Microbiol 75:5972–5980. CrossRefPubMedPubMedCentralGoogle Scholar
  6. Chen W, Chen H, Xia Y, Yang J, Zhao J, Tian F, Zhang HP, Zhang H (2009) Immobilization of recombinant thermostable β-galactosidase from Bacillus stearothermophilus for lactose hydrolysis in milk. J Dairy Sci 92:491–498. CrossRefPubMedGoogle Scholar
  7. Du ZJ, Lv GQ, Rooney AP, Miao TT, Xu QQ, Chen GJ (2011) Agarivorans gilvus sp. nov. isolated from seaweed. Int J Syst Evol Microbiol 61:493–496. CrossRefPubMedGoogle Scholar
  8. Dumortier V, Brassart C, Bouquelet S (1994) Purification and properties of a β-D-galactosidase from Bifidobacterium Bifidum exhibiting a transgalactosyl reaction. Biotechnol Appl Biochem 19:341–354Google Scholar
  9. Erich S, Kuschel B, Schwarz T, Ewert J, Böhmer N, Niehaus F, Eck J, Lutz-Wahl S, Stressler T, Fischer L (2015) Novel high-performance metagenome β-galactosidases for lactose hydrolysis in the dairy industry. J Biotechnol 210:27–37. CrossRefPubMedGoogle Scholar
  10. Gatt S, Baker EA (1970) Purification and separation of α- and β-galactosidases from spinach leaves. Biochim Biophys Acta 206:125–135. CrossRefPubMedGoogle Scholar
  11. Ha SC, Lee S, Lee J, Kim HT, Ko HJ, Kim KH, Choi IG (2011) Crystal structure of a key enzyme in the agarolytic pathway, α-neoagarobiose hydrolase from Saccharophagus degradans 2-40. Biochem Biophys Res Commun 412:238–244. CrossRefPubMedGoogle Scholar
  12. Haider T, Husain Q (2007) Calcium alginate entrapped preparations of Aspergillus oryzae β galactosidase: its stability and applications in the hydrolysis of lactose. Int J Biol Macromol 41:72–80. CrossRefPubMedGoogle Scholar
  13. Hatada Y, Ohta Y, Horikoshi K (2006) Hyperproduction and application of α-agarase to enzymatic enhancement of antioxidant activity of porphyran. J Agric Food Chem 54:9895–9900. CrossRefPubMedGoogle Scholar
  14. Henrissat B (1991) A classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem J 280:309–316. CrossRefPubMedPubMedCentralGoogle Scholar
  15. Hidaka M, Fushinobu S, Ohtsu N, Motoshima H, Matsuzawa H, Shoun H, Wakagi T (2002) Trimeric crystal structure of the glycoside hydrolase family 42 β-galactosidase from Thermus thermophilus A4 and the structure of its complex with galactose. J Mol Biol 322:79–91. CrossRefPubMedGoogle Scholar
  16. Hsu CA, Lee SL, Chou CC (2007) Enzymatic production of galactooligosaccharides by β-galactosidase from Bifidobacterium longum BCRC 15708. J Agric Food Chem 55:2225–2230. CrossRefPubMedGoogle Scholar
  17. Husain Q (2010) β Galactosidases and their potential applications: a review. Crit Rev Biotechnol 30:41–62. CrossRefPubMedGoogle Scholar
  18. Jung S, Lee CR, Chi WJ, Bae CH, Hong SK (2017) Biochemical characterization of a novel cold-adapted GH39 β-agarase, AgaJ9, from an agar-degrading marine bacterium Gayadomonas joobiniege G7. Appl Microbiol Biotechnol 101:1965–1974. CrossRefPubMedGoogle Scholar
  19. Kishore D, Kayastha AM (2012) A β-galactosidase from chick pea (Cicer arietinum) seeds: its purification, biochemical properties and industrial applications. Food Chem 134:1113–1122. CrossRefPubMedGoogle Scholar
  20. Kumar S, Tamura K, Nei M (1994) MEGA: molecular evolutionary genetics analysis software for microcomputers. Comput Appl Biosci 10:189–191. PubMedCrossRefGoogle Scholar
  21. Lee CH, Kim HT, Yun EJ, Lee AR, Kim SR, Kim JH, Choi IG, Kim KH (2014) A novel agarolytic β-galactosidase acts on agarooligosaccharides for complete hydrolysis of agarose into monomers. Appl Environ Microbiol 80:5965–5973. CrossRefPubMedPubMedCentralGoogle Scholar
  22. Liang YX, Ma XQ, Zhang LJ, Li FL, Liu Z, Mao XZ (2017) Biochemical characterization and substrate degradation mode of a novel exo-type β-agarase from Agarivorans gilvus WH0801. J Agric Food Chem 65:7982–7988. CrossRefPubMedGoogle Scholar
  23. Lin B, Liu Y, Lu GY, Zhao M, Hu Z (2017) An agarase of glycoside hydrolase family 16 from marine bacterium Aquimarina agarilytica ZC1. FEMS Microbiol Lett 364.
  24. Liu N, Mao XZ, Du ZJ, Mu BZ, Wei DZ (2014a) Cloning and characterisation of a novel neoagarotetraose-forming-β-agarase, AgWH50A from Agarivorans gilvus WH0801. Carbohydr Res 388:147–151. CrossRefPubMedGoogle Scholar
  25. Liu N, Mao XZ, Meng Y, Mu BZ, Wei DZ (2014b) Gene cloning, expression and characterisation of a new β-agarase, AgWH50C, producing neoagarobiose from Agarivorans gilvus WH0801. World J Microbiol Biotechnol 30:1691–1698. CrossRefPubMedGoogle Scholar
  26. Liu N, Yang M, Mao XZ, Mu BZ, Wei DZ (2016) Molecular cloning and expression of a new α-neoagarobiose hydrolase from Agarivorans gilvus WH0801 and enzymatic production of 3,6-anhydro-L-galactose. Biotechnol Appl Biochem 63:230–237. CrossRefPubMedGoogle Scholar
  27. Nath A, Mondal S, Chakraborty S, Bhattacharjee C, Chowdhury R (2014) Production, purification, characterization, immobilization, and application of β-galactosidase: a review. Asia Pac J Chem Eng 9:330–348. CrossRefGoogle Scholar
  28. Niu DD, Tian XJ, Mchunu NP, Jia C, Singh S, Liu XG, Prior BA, Lu FP (2017) Biochemical characterization of three Aspergillus niger β-galactosidases. Electron J Biotechnol 27:37–43. CrossRefGoogle Scholar
  29. Oliveira C, Guimarães PM, Domingues L (2011) Recombinant microbial systems for improved β-galactosidase production and biotechnological applications. Biotechnol Adv 29:600–609. CrossRefPubMedGoogle Scholar
  30. Onishi N, Tanaka T (1995) Purification and properties of a novel thermostable galacto-oligosaccharide-producing β-galactosidase from Sterigmatomyces elviae CBS8119. Appl Environ Microbiol 61:4026–4030 0099–2240/95/$04.00+0PubMedPubMedCentralGoogle Scholar
  31. Pawlak-Szukalska A, Wanarska M, Popinigis AT, Kur J (2014) A novel cold-active β-D-galactosidase with transglycosylation activity from the Antarctic Arthrobacter sp. 32cB—gene cloning, purification and characterization. Process Biochem 49:2122–2133. CrossRefGoogle Scholar
  32. Rees DA (1969) Structure, conformation, and mechanism in the formation of polysaccharide gels and networks. Adv Carbohydr Chem Biochem 24:267–332. CrossRefPubMedGoogle Scholar
  33. Rosenberg AH, Studier FW (1987) T7 RNA polymerase can direct expression of influenza virus cap-binding protein (PB2) in Escherichia coli. Gene 59:191–200. CrossRefPubMedGoogle Scholar
  34. Sakai T, Tsuji H, Shibata S, Hayakawa K, Matsumoto K (2008) Repeated-batch production of galactooligosaccharides from lactose at high concentration by using alginate-immobilized cells of Sporobolomyces singularis YIT 10047. Appl Microbiol 54:285–293. CrossRefGoogle Scholar
  35. Seok JH, Kim HS, Hatada Y, Nam SW, Kim YH (2012) Construction of an expression system for the secretory production of recombinant α-agarase in yeast. Biotechnol Lett 34:1041–1049. CrossRefPubMedGoogle Scholar
  36. Studier FW, Rosenberg AH, Dunn JJ, Dubendorff JW (1990) Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol 185:60–89CrossRefPubMedGoogle Scholar
  37. Syazni, Yanagisawa M, Kasuu M, Ariga O, Nakasaki K (2015) Direct production of ethanol from neoagarobiose using recombinant yeast that secretes α-neoagarooligosaccharide hydrolase. Enzyme Microb Technol 85:82–89. CrossRefPubMedGoogle Scholar
  38. Walton A, Smith VJ (1999) Primary culture of the hyaline haemocytes from marine decapods. Fish Shellfish Immunol 9:181–194. CrossRefGoogle Scholar
  39. Yun JE, Choi I, Kim HK (2015) Red macroalgae as a sustainable resource for bio-based products. Trends Biotechnol 33:247–249. CrossRefPubMedGoogle Scholar
  40. Zhang PJ, Rui JP, Du ZJ, Xue CH, Li XZ, Mao XZ (2016) Complete genome sequence of Agarivorans gilvus WH0801T, an agarase-producing bacterium isolated from seaweed. J Biotechnol 219:22–23. CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.College of Food Science and EngineeringOcean University of ChinaQingdaoChina
  2. 2.Laboratory for Marine Drugs and BioproductsQingdao National Laboratory for Marine Science and TechnologyQingdaoChina

Personalised recommendations