β-Glucanase specific expression in the intestine of transgenic pigs

  • Li-zeng Guan
  • Shuai Zhao
  • Gang Shu
  • Qing-yan Jiang
  • Geng-yuan Cai
  • Zhen-fang Wu
  • Qian-yun XiEmail author
  • Yong-liang ZhangEmail author
Original Paper


Producing heterologous enzymes in the animal digestive tract to improve feed utilization rate is a new research strategy by transgenic technology. In this study, transgenic pigs specifically expressing β-glucanase gene in the intestine were successfully produced by somatic cell nuclear transfer technology in order to improve digestibility of dietary β-glucan and absorption of nutrients. The β-glucanase activity in the intestinal juice of 4 transgenic pigs was found to be 8.59 ± 2.49 U/mL. The feeding trial results showed that the crude protein digestion of 4 transgenic pigs was significantly increased compared with that of the non-transgenic pigs. In order to investigate the inheritance of the transgene, 7 G1 transgenic pigs were successfully obtained. The β-glucanase activity in the intestinal juice of 7 G1 transgenic pigs was found to be 2.35 ± 0.72 U/mL. The feeding trial results showed the crude protein digestion and crude fat digestion were significantly higher in 7 G1 transgenic pigs than in non-transgenic pigs. Taken together, our study demonstrated that the foreign β-glucanase expressing in the intestine of the transgenic pigs could reduce the anti-nutritional effect of β-glucans in feed. In addition, β-glucanase gene could be inherited to the offsprings and maintain its physiological function. It is a promising approach to improve feed utilization by producing transgenic animals.


β-Glucanase Intestine Transgenic pigs 



This work was supported by grants from the Key Project of Transgenic Animal (2014ZX0800948B).

Compliance with ethical standards

Conflict of interest

None of the authors declare any conflicts of interest.


  1. Bensadoun A, Weinstein D (1976) Assay of proteins in the presence of interfering materials. Anal Biochem 70(1):241–250CrossRefGoogle Scholar
  2. Buckow R, Heinz V, Knorr D (2015) Effect of high hydrostatic pressure–temperature combinations on the activity of β-glucanase from barley malt. J Inst Brew 111(3):282–289CrossRefGoogle Scholar
  3. Budiani A, Putranto RA, Minarsih H, Fitranti N, Santoso D (2009) Rapid cloning for gene encoding fungal β-1,6-glucanase by means of RT-PCR using specific primers. Menara Perkeb 77(1):47–57Google Scholar
  4. Chang SK, Dohrman AF, Basbaum CB, Ho SB, Tsuda T, Toribara NW, Gum JR, Kim YS (1994) Localization of mucin (MUC2 and MUC3) messenger RNA and peptide expression in human normal intestine and colon cancer. Gastroenterology 107(1):28–36CrossRefGoogle Scholar
  5. Cranston A, Dong C, Howcroft J, Clark AJ (2001) Chromosomal sequences flanking an efficiently expressed transgene dramatically enhance its expression. Gene 269(1):217–225CrossRefGoogle Scholar
  6. Eszterhas SK, Bouhassira EE, Martin DIK, Fiering S (2002) Transcriptional interference by independently regulated genes occurs in any relative arrangement of the genes and is influenced by chromosomal integration position. Mol Cell Biol 22(2):469–479CrossRefGoogle Scholar
  7. Fontes CM, Ali S, Gilbert HJ, Hazlewood GP, Hirst BH, Hall J (1999) Bacterial xylanase expression in mammalian cells and transgenic mice. J Biotechnol 72(1–2):95–101CrossRefGoogle Scholar
  8. Guan LZ, Sun YP, Xi QY, Wang JL, Zhou JY, Shu G, Jiang QY, Zhang YL (2013) β-Glucanase specific expression in the parotid gland of transgenic mice. Transgenic Res 22(4):805–812CrossRefGoogle Scholar
  9. Guan L, Xi Q, Sun Y, Wang J, Zhou J, Shu G, Jiang Q, Zhang Y (2014) Intestine-specific expression of the β-glucanase in mice. Can J Anim Sci 94(2):287–293CrossRefGoogle Scholar
  10. Guan LZ, Cai JS, Zhao S, Sun YP, Wang JL, Jiang Y, Shu G, Jiang QY, Wu ZF, Xi QY (2016) Improvement of anti-nutritional effect resulting from β-glucanase specific expression in the parotid gland of transgenic pigs. Transgenic Res 26(1):1–11CrossRefGoogle Scholar
  11. Gum Jr., Hicks JW, Gillespie AM, Carlson EJ, Kömüves L, Karnik S, Hong JC, Epstein CJ, Kim YS (1999) Goblet cell-specific expression mediated by the MUC2 mucin gene promoter in the intestine of transgenic mice. Am J Physiol 276(3 Pt 1):G666Google Scholar
  12. Lai L, Prather RS (2003) Production of cloned pigs by using somatic cells as donors. Cloning Stem Cells 5(4):233CrossRefGoogle Scholar
  13. Lv YM, Tan WP, Xiao CL, Fan MR, Liao YL (2014) Effect of high temperature on starch formation of grain and activities of enzymes related to starch synthesis of quality rice varieties. Acta Agriculturae Boreali-Sinica 29(1):135–139Google Scholar
  14. Melo EO, Canavessi AM, Franco MM, Rumpf R (2007) Animal transgenesis: state of the art and applications. J Appl Genet 48:47–61CrossRefGoogle Scholar
  15. Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Biochem 31(3):426–428Google Scholar
  16. Murphy P, Bello FD, O’Doherty JV, Arendt EK, Sweeney T, Coffey A (2012) Effects of cereal β-glucans and enzyme inclusion on the porcine gastrointestinal tract microbiota. Anaerobe 18(6):557–565CrossRefGoogle Scholar
  17. Nakanishi T et al (2002) FISH analysis of 142 EGFP transgene integration sites into the mouse genome. Genomics 80:564–574CrossRefGoogle Scholar
  18. Pei H, Guo X, Yang W, Lv J, Chen Y, Cao Y (2015) Directed evolution of a β-1,3-1,4-glucanase from Bacillus subtilis MA139 for improving thermal stability and other characteristics. J Basic Microbiol 55(7):869–878CrossRefGoogle Scholar
  19. Ravindran V, Tilman ZV, Pch M, Ravindran G, Coles GD (2007) Influence of β-glucanase supplementation on the metabolisable energy and ileal nutrient digestibility of normal starch and waxy barleys for broiler chickens. Anim Feed Sci Technol 134(1):45–55CrossRefGoogle Scholar
  20. Tytgat KM, Büller HA, Opdam FJ, Kim YS, Einerhand AW, Dekker J (1994) Biosynthesis of human colonic mucin: Muc2 is the prominent secretory mucin. Gastroenterology 107(5):1352CrossRefGoogle Scholar
  21. Walsh GA, Headon DR (1993) Enzymes in the animal-feed industry. Trends Biotechnol 11(10):424–430CrossRefGoogle Scholar
  22. Woyewoda AD, Shaw S, Ke PJ, Burns BG (1986) Recommended laboratory methods for assessment of fish quality. Can Tech Rep Fish Aquat Sci 143:1448Google Scholar
  23. Yin Z, Kong QR, Zhao ZP, Wu ML, Mu YS, Hu K, Liu ZH (2012) Position effect variegation and epigenetic modification of a transgene in a pig model. Genetics Mol Res GMR 11(1):355CrossRefGoogle Scholar
  24. Zhang XW, Li ZC, Yang HQ, Liu DW, Cai GY, Li GL, Mo JX, Wang DH, Zhong CL, Wang HQ, Sun Y, Shi JS, Zheng EQ, Meng FM, Zhang M, He XY, Zhou R, Zhang J, Huang MR, Zhang R, Li N, Fan MZ, Yang JZ, Wu ZF (2018) Novel transgenic pigs with enhanced growth and reduced environmental impact. eLife 7:e34286CrossRefGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  • Li-zeng Guan
    • 1
  • Shuai Zhao
    • 2
  • Gang Shu
    • 2
  • Qing-yan Jiang
    • 2
  • Geng-yuan Cai
    • 2
  • Zhen-fang Wu
    • 2
  • Qian-yun Xi
    • 2
    Email author
  • Yong-liang Zhang
    • 2
    Email author
  1. 1.College of Agriculture and Forestry ScienceLinyi UniversityLinyi CityChina
  2. 2.National Engineering Research Center for Breeding Swine Industry, Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding, College of Animal Science, SCAU-Alltech Research Joint AllianceSouth China Agricultural UniversityGuangzhouChina

Personalised recommendations