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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2018, Vol. 38 Issue (6): 43-51    DOI: 10.13523/j.cb.20180606
    
Identification of Cyclic β-1,2-glucan Produced under pH Control
Juan-juan QU,Xiao-bei ZHAN(),Hong-tao ZHANG(),Xian-chao ZHOU,Yu-chen JIA,Xue-ying CAO
Key Laboratory of Carbohydrate Chemistry and Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122, China
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Abstract  

Objective: To study the effects of pH on the production cyclic β-1,2-glucan, and analyze the structure of cyclic glucans under pH control. Methods:Rhizobium radiobacter ATCC 1333 was used to analyze the effects of pH on fermentation. The cyclic glucans were separated, purified and purified, then analyzed by matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-MS), monosaccharide composition analysis, electrospray ionization mass spectrometry(ESI-MS), fourier transform infrared spectrometer(FTIR) and nuclear magnetic resonance spectroscopy(NMR). Results: The two stage pH strategy was developed in which pH was controlled at 7.0 for the cell growth and then shift to 5.5. The results showed that biomass was increased by 102% and the cyclic glucan concentration increased by 52%, and the fermentation color was controlled at the background level. Also, the glucans from Rhizobium radiobacter ATCC 1333 were cyclic glucans consisting of glucose by β-1,2 linkages with a degree of polymerization ranging from 17~22, mainly 19. Conclusion: The pH controlled at 7.0 has no effect on the structure of cyclic β-1,2-glucans, and provides a theoretical basis for the fermentation optimization and reliable source of cyclic β-1,2-glucans.

Key words： Rhizobium radiobacter ATCC 1333      pH control Cyclic      β-1,2-glucan      MALDI-MS      ESI-MS      NMR     
Received: 07 February 2018      Published: 06 July 2018
ZTFLH:  Q815  
Corresponding Authors: Xiao-bei ZHAN,Hong-tao ZHANG     E-mail: xbzhan@yahoo.com;htzhang@jiangnan.edu.cn
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Juan-juan QU
Xiao-bei ZHAN
Hong-tao ZHANG
Xian-chao ZHOU
Yu-chen JIA
Xue-ying CAO
Cite this article:

Juan-juan QU,Xiao-bei ZHAN,Hong-tao ZHANG,Xian-chao ZHOU,Yu-chen JIA,Xue-ying CAO. Identification of Cyclic β-1,2-glucan Produced under pH Control. China Biotechnology, 2018, 38(6): 43-51.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20180606     OR     https://manu60.magtech.com.cn/biotech/Y2018/V38/I6/43

Fig.1 The effects of initial pH on fermentation process
Fig.2 The full-wavelength scanning of the color of the fermentation broth at different fermentation time
Fig.3 The batch profile of cyclic glucans
Fig.4 The batch profile of cell growth and cyclic glucan production under pH control
pH Maximal biomass
/(g/L)		 Final glucans production
/(g/L)		 Yp/x/(g/g)
Cell growth stage Production stage
7.0 uncontrol 6.24 2.24 0.358
6.0 6.32 2.51 0.397
5.5 6.36 2.79 0.439
5.0 6.19 2.6 0.420
Table 1 Effect of different pH controlled on batch fermentation of cyclic glucans
Fig.5 The Monosaccharide compositions analysis (a)The target glucans (b) The standards (from 1 to 9: glucosamine, rhamnose, arabinose, galactosamine, galactose, glucose, xylose, mannose and fructose)
Fig.6 The MALDI-MS identification of the glucans
Observed mass m/z
[M+Na]+		 Theor. Mass Assignment
2 777.8 2 754 CβG17
2 939.9 2 916 CβG18
3 101.9 3 078 CβG19
3 264.0 3 240 CβG20
3 426.0 3 402 CβG21
3 589.1 3 564 CβG22
Table 2 Analysis of MALDI-MS data of the fermentation broth
Fig.7 ESI-CID-MS/MS analysis of hydrolyzed target glucans
Fig.8 The FTIR spectra of the glucans
Fig.9 NMR spectroscopy of glucans from Rhizobium radiobacter NBRC13259 (a) 1H-NMR (b) 13C-NMR (c) The structure of cyclic β-1,2-glucans
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