Optimized Fermentation for Production of Xanthan Gum Using Xanthomonas axonopodis
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摘要: 为了获得更高品质的黄原胶，优化黄原胶高产菌株地毯草黄单胞菌FJAT-10151的发酵工艺，通过单因素及正交设计对培养基（碳源、氮源和无机盐离子）和发酵条件（发酵时间、pH、装液量和接种量）进行优化，试验获得最佳发酵培养基为葡萄糖30 g·L-1、豆饼粉30 g·L-1、KH2PO42 g·L-1、pH值9.0。在装液量50 mL/250 mL、接种量8%培养条件下发酵72 h，黄原胶产量达到21.0 g·L-1，是初始培养条件产量8.65 g·L-1的2.43倍。优化发酵工艺后，黄原胶品质提高，丙酮酸含量从8.9%升高到16.3%，而蛋白质含量从15.27%降低到4.8%。Abstract: Conditions of fermentation to produce extracellular xanthan gum by using Xanthomonas axonopodis FJAT-10151 were optimized to maximize quality of the gum. Single factor and orthogonal experiments on medium composition (including sources of carbon, nitrogen and inorganic ions) and fermentation conditions (including pH, time duration, fill volume and inoculum ratio) were conducted. It was found that using a medium consisting of glucose 30 g·L-1, soybean cake powder 30 g·L-1, and KH2PO4 2 g·L-1 at pH 9.0 with a filled culture of 50 mL containing 8% of inoculum per 250 mL flask, a yield of 21.0 g·L-1 of xanthan gum could be obtained after fermentation for 72 h. The production capacity was 243% higher than that prior to the optimization. More importantly, a higher quality of xanthan gum with the content of pyruvic acid increased from 8.9% to 16.3% and the content of protein decreased from 15.27% to 4.8% was realized.
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Key words:
- Xanthomonas axonopodis /
- xanthan gum /
- orthogonal design
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图 1 黄原胶产量随碳源的变化
注：图上不同字母表示经Duncan法检测在0.05水平上差异显著，下同。
Figure 1. Change on xanthan gum production by varying carbon source
图 3 黄原胶产量随无机盐离子的变化
Figure 3. Change on xanthan gum production by varying inorganic salt addition
图 4 黄原胶产量随发酵时间的变化
Figure 4. Change on xanthan gum production by varying fermentation 18:13:09
图 5 黄原胶产量随装液量的变化
Figure 5. Change on xanthan gum production due to filled culture volume
图 6 黄原胶产量随初始pH的变化
Figure 6. Change on xanthan gum production due to initial pH of medium
图 8 地毯草黄单胞菌FJAT-10151的透射电镜观察
Figure 8. Electron microscopy of X. axonopodis FJAT-10151
表 1 L9(34)正交试验分析因素与水平
Table 1. Factors and levels of orthogonal experiment
因素 因素/(g·L-1) 葡萄糖 豆饼粉 KH2PO4 -1 20 10 1 0 30 20 2 1 40 30 4 
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表 2 正交优化结果极差分析
Table 2. Range analysis on orthogonal test results
水平 因素/(g·L-1) 黄原胶产量
/(g·L-1)葡萄糖 豆饼粉 KH2PO4 1 40 30 1 15.29±1.83 2 20 20 4 13.61±1.22 3 40 10 4 11.14±0.55 4 20 30 2 14.39±0.48 5 30 30 4 17.71±1.02 6 40 20 2 16.34±0.19 7 30 20 1 13.83±0.43 8 30 10 2 12.07±0.37 9 20 10 1 9.51±0.09 低 20 10 1 - 中 30 20 2 - 高 40 30 4 - Ⅰ 0.625 0.545 0.644 - Ⅱ 0.713 0.730 0.713 - Ⅲ 0.727 0.790 0.708 - R 0.102 0.245 0.070 -
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表 3 黄原胶中中性糖、丙酮酸和蛋白质的含量
Table 3. Neutral sugar, pyruvic acid and protein contents in xanthan gum produced from fermentation
项目 中性糖
/%丙酮酸
/%蛋白质
/%标准曲线 葡萄糖 葡萄糖醛酸 牛血清白蛋白 相关系数(R2) 0.9980 0.9982 0.9980 黄原胶1 36.81 8.9 15.27 黄原胶2 25.3 16.3 4.8 注：黄原胶1：基础培养基提取的黄原胶[10]；黄原胶2：优化培养基后提取的黄原胶。中性糖线性方程为OD490nm=0.0331CGlu +0.1151；丙酮酸线性方程为OD530nm=15.256CGlcA+0.0782；蛋白质线性方程为OD580nm=0.0293CBSA +0.4233。
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