利用退浆废水发酵生产脂肽及其在染整工艺上的应用
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摘要
微生物代谢产生的脂肽类生物表面活性剂具有表面活性强、生物降解性好等特性,在化妆品、环境、医药、食品和纺织等领域呈现出潜在的应用价值。然而,脂肽类生物表面活性剂的产量低,分离纯化困难,生产成本高,在很大程度上限制了其广泛的应用。本论文以产生脂肽的枯草芽孢杆菌(Bacillus subtilis HSO121)为出发菌株,利用染整工业的退浆废水作为枯草芽孢杆菌生长的碳源,配制成发酵培养基发酵生产脂肽。然后通过减少分离纯化步骤的方法得到脂肽粗品,并探索了其在各染整工艺上的应用。
取商用的淀粉浆料作为菌种生长的碳源,以菌浓、脂肽产量和无菌发酵液表面张力为评价参数,采用单因素和正交实验的方法对Bacillus subtilis HSO121产脂肽的培养条件进行优化,在最优培养基上培养,脂肽产量由初始的0.67 g/l提高到1.24 g/l。
研究了退浆废水中各主要成分对脂肽产量的影响,结果表明培养基中NaCl在10g/l以下时对菌种生长代谢产生促进作用;培养基中渗透剂JFC不超过0.1 g/l时也会对脂肽的代谢产生促进作用;浆料中PVA少量的存在,不会对菌体的生长代谢产生明显的影响,但PVA应占浆料的30%以下。
取预处理好的退浆废水按照最优培养基配方配制成发酵培养基。菌种在退浆废水培养基中生长良好,脂肽产量达到了1.03g/l。此外,退浆废水培养基在发酵前其COD为41360 mg/l,发酵后废液的COD为22880 mg/l,COD值降低了近一半。
在退浆废水培养基中发酵产生的脂肽经分离提纯后,薄层层析和红外光谱的实验结果表明该发酵产物为环状的脂肽类生物表面活性剂。产物带电性检测结果为阴离子表面活性剂。产物对液体石蜡的乳化能力很强,24小时后的乳化指数为83%,且具有很高的乳化稳定性。该脂肽具有很低的临界胶束浓度,其CMC值为50 mg/l,此时的表面张力值为28.53 mN/m。
经活性炭脱色后的脂肽粗制品作为活性染料染色织物皂洗剂的实验中。皂洗的试样为50%水解率的活性黄3RS和活性蓝BRF染色后的府绸织物,得到了pH值为9,脂肽浓度为0.05g/l,皂洗温度为75℃的最佳皂洗工艺条件。脂肽类生物表面活性剂对活性红3BS,活性黄3RS,活性蓝BRF染色的织物都具有很好的皂洗效果。与皂片皂洗效果相比,脂肽去除浮色的能力更强,湿摩擦牢度提高了近1个等级,而且脂肽皂洗所需温度更低,用量更少,更加地节能环保。
脂肽粗制品作为织物抗菌剂的实验中,利用抑菌圈法和振荡法对脂肽整理后织物抗菌性能进行测试。抑菌圈法中,经抗菌整理的织物对于金黄色葡萄球菌和大肠杆菌的抑菌圈宽度分别为1.4 mm和2.1 mm,两个抑菌圈宽度D＞1 mm,说明脂肽整理的织物为溶出型抗菌织物,且抗菌效果好。振荡法实验中,对金黄色葡萄球菌的抑菌率为82.17%,对大肠杆菌的抑菌率为84.02%,说明织物经脂肽整理后,具有很好的抗菌效果。
脂肽作为羊毛织物的柔软整理剂实验中,没有经脂肽柔软整理的空白羊毛织物的弯曲长度为18.9 mm,经过脂肽柔软整理的羊毛织物的弯曲长度为13.7 mm。这表明了经脂肽柔软整理后羊毛织物的弯曲长度有了明显的降低,羊毛织物的柔软性能得到了改善。
Lipopeptides have gained more and more attention due to their excellent surface activities, biodegradability, low toxicity, and environmentally friendly. Lipopeptides exhibited potential applications in cosmetic, environmental fields, biomedicine, food and textile industry. However, its applications have been restricted to a great extent because of their low yields and complex purification procedures and high costs. In this dissertation, we reported the production of lipopeptides using desizing wastewater as alternative carbon source by Bacillus subtilis HSO121, and discussed the applications of crude lipopeptides in Dyeing & Finishing technology after simplifying purification procedures.
In this paper, we used the commercial starch as carbon source. Single factor test and orthogonal experiment were used to optimize the production of lipopeptides by Bacillus subtilis HSO121. The biomass, lipopeptides yields and reduced surface tension values were selected as evaluating parameters. Grown in the optimized medium, lipopeptides yields improved from 0.67 g/L to 1.24 g/L.
The effects of main composition in the desizing wastewater on lipopeptides yields were investigated. The results demonstrated that the concentration of NaCl less than 10g/l and the concentration of penetrant JFC less than 0.1 g/l in medium could contribute to growth and metabolism of bacteria, a small amount of PVA had no significant effect on growth and metabolism of bacteria, but PVA should less than 30% in sizing agent.
Bacillus subtilis HSO121 grew well in the desizing wastewater medium, which prepared by pretreated desizing wastewater according to the optimized medium, the yields of lipopeptides attained 1.03 g/L. Moreover, the Chemical Oxygen Demand of the desizing wastewater after cultivation reduced by half compared with before cultivation, decreased from 41360 mg/L to 22880 mg/L.
The lipopeptides were separated and extracted from the desizing wastewater medium after cultivation, and characterized by thin layer chromatography (TLC) on silica gel and infrared ray (IR) analysis. The charged detection for lipopeptides was anionic-surfactant. Emulsification activity of the lipopeptides from Bacillus subtilis HSO121 was measured with water-immiscible substrates. The results show that the lipopeptides had high emulsification activity and emulsification stability against liquid paraffin. After 24 hours the emulsification index was 83%. The CMC of the lipopeptides was approximately 50 mg/L, and the lipopeptides reduced the surface tension of the water from 72 to 28.53mN/m.
In order to reduce the cost used, the crude lipopeptides were prepared by discoloring. The crude lipopeptides as soaping agents in soaping process were investigated. The test sample was poplin dyed with partially hydrolyzed reactive red 3BS, reactive yellow 3RS and reactive blue BRF. The optimal soaping process was obtained: pH=9, 0.05g/L of lipopeptides, at 75°C. Regardless of dyeing by partially hydrolyzed reactive red 3BS, reactive yellow 3RS or reactive blue BRF, the results of soaping with lipopeptides were better than traditional soaping process. Compared with traditional soaping process, lipopeptides soaping showed higher efficient, higher colorfastness, more energy-efficient and more environmentally friendly.
The crude Lipopeptides as fabric antibacterial agents were investigated. In the agar diffusion plate method, the fabric after antibacterial finish showed a large and clear inhibition zone on solid agar medium against E. coli and S. aureus, the width of the inhibition zone was 2.1 mm and 1.4 mm respectively. It indicated that the fabric after antibacterial finish was leaching type antibacterial fabric, and had good antibacterial effect. In the shake flask method, the finished fabric showed over 84.02% bacterial reduction against E. coli, and over 82.17% bacterial reduction against S. aureus. It is well recognized that lipopeptides has good antimicrobial activity.
The crude Lipopeptides as fabric softener were investigated. The bending length of wool fabric before softening finishes was 18.9 mm and after softening finishes was 13.7 mm. The results showed that lipopeptides as softener improved performance of wool soft.
取商用的淀粉浆料作为菌种生长的碳源,以菌浓、脂肽产量和无菌发酵液表面张力为评价参数,采用单因素和正交实验的方法对Bacillus subtilis HSO121产脂肽的培养条件进行优化,在最优培养基上培养,脂肽产量由初始的0.67 g/l提高到1.24 g/l。
研究了退浆废水中各主要成分对脂肽产量的影响,结果表明培养基中NaCl在10g/l以下时对菌种生长代谢产生促进作用;培养基中渗透剂JFC不超过0.1 g/l时也会对脂肽的代谢产生促进作用;浆料中PVA少量的存在,不会对菌体的生长代谢产生明显的影响,但PVA应占浆料的30%以下。
取预处理好的退浆废水按照最优培养基配方配制成发酵培养基。菌种在退浆废水培养基中生长良好,脂肽产量达到了1.03g/l。此外,退浆废水培养基在发酵前其COD为41360 mg/l,发酵后废液的COD为22880 mg/l,COD值降低了近一半。
在退浆废水培养基中发酵产生的脂肽经分离提纯后,薄层层析和红外光谱的实验结果表明该发酵产物为环状的脂肽类生物表面活性剂。产物带电性检测结果为阴离子表面活性剂。产物对液体石蜡的乳化能力很强,24小时后的乳化指数为83%,且具有很高的乳化稳定性。该脂肽具有很低的临界胶束浓度,其CMC值为50 mg/l,此时的表面张力值为28.53 mN/m。
经活性炭脱色后的脂肽粗制品作为活性染料染色织物皂洗剂的实验中。皂洗的试样为50%水解率的活性黄3RS和活性蓝BRF染色后的府绸织物,得到了pH值为9,脂肽浓度为0.05g/l,皂洗温度为75℃的最佳皂洗工艺条件。脂肽类生物表面活性剂对活性红3BS,活性黄3RS,活性蓝BRF染色的织物都具有很好的皂洗效果。与皂片皂洗效果相比,脂肽去除浮色的能力更强,湿摩擦牢度提高了近1个等级,而且脂肽皂洗所需温度更低,用量更少,更加地节能环保。
脂肽粗制品作为织物抗菌剂的实验中,利用抑菌圈法和振荡法对脂肽整理后织物抗菌性能进行测试。抑菌圈法中,经抗菌整理的织物对于金黄色葡萄球菌和大肠杆菌的抑菌圈宽度分别为1.4 mm和2.1 mm,两个抑菌圈宽度D＞1 mm,说明脂肽整理的织物为溶出型抗菌织物,且抗菌效果好。振荡法实验中,对金黄色葡萄球菌的抑菌率为82.17%,对大肠杆菌的抑菌率为84.02%,说明织物经脂肽整理后,具有很好的抗菌效果。
脂肽作为羊毛织物的柔软整理剂实验中,没有经脂肽柔软整理的空白羊毛织物的弯曲长度为18.9 mm,经过脂肽柔软整理的羊毛织物的弯曲长度为13.7 mm。这表明了经脂肽柔软整理后羊毛织物的弯曲长度有了明显的降低,羊毛织物的柔软性能得到了改善。
Lipopeptides have gained more and more attention due to their excellent surface activities, biodegradability, low toxicity, and environmentally friendly. Lipopeptides exhibited potential applications in cosmetic, environmental fields, biomedicine, food and textile industry. However, its applications have been restricted to a great extent because of their low yields and complex purification procedures and high costs. In this dissertation, we reported the production of lipopeptides using desizing wastewater as alternative carbon source by Bacillus subtilis HSO121, and discussed the applications of crude lipopeptides in Dyeing & Finishing technology after simplifying purification procedures.
In this paper, we used the commercial starch as carbon source. Single factor test and orthogonal experiment were used to optimize the production of lipopeptides by Bacillus subtilis HSO121. The biomass, lipopeptides yields and reduced surface tension values were selected as evaluating parameters. Grown in the optimized medium, lipopeptides yields improved from 0.67 g/L to 1.24 g/L.
The effects of main composition in the desizing wastewater on lipopeptides yields were investigated. The results demonstrated that the concentration of NaCl less than 10g/l and the concentration of penetrant JFC less than 0.1 g/l in medium could contribute to growth and metabolism of bacteria, a small amount of PVA had no significant effect on growth and metabolism of bacteria, but PVA should less than 30% in sizing agent.
Bacillus subtilis HSO121 grew well in the desizing wastewater medium, which prepared by pretreated desizing wastewater according to the optimized medium, the yields of lipopeptides attained 1.03 g/L. Moreover, the Chemical Oxygen Demand of the desizing wastewater after cultivation reduced by half compared with before cultivation, decreased from 41360 mg/L to 22880 mg/L.
The lipopeptides were separated and extracted from the desizing wastewater medium after cultivation, and characterized by thin layer chromatography (TLC) on silica gel and infrared ray (IR) analysis. The charged detection for lipopeptides was anionic-surfactant. Emulsification activity of the lipopeptides from Bacillus subtilis HSO121 was measured with water-immiscible substrates. The results show that the lipopeptides had high emulsification activity and emulsification stability against liquid paraffin. After 24 hours the emulsification index was 83%. The CMC of the lipopeptides was approximately 50 mg/L, and the lipopeptides reduced the surface tension of the water from 72 to 28.53mN/m.
In order to reduce the cost used, the crude lipopeptides were prepared by discoloring. The crude lipopeptides as soaping agents in soaping process were investigated. The test sample was poplin dyed with partially hydrolyzed reactive red 3BS, reactive yellow 3RS and reactive blue BRF. The optimal soaping process was obtained: pH=9, 0.05g/L of lipopeptides, at 75°C. Regardless of dyeing by partially hydrolyzed reactive red 3BS, reactive yellow 3RS or reactive blue BRF, the results of soaping with lipopeptides were better than traditional soaping process. Compared with traditional soaping process, lipopeptides soaping showed higher efficient, higher colorfastness, more energy-efficient and more environmentally friendly.
The crude Lipopeptides as fabric antibacterial agents were investigated. In the agar diffusion plate method, the fabric after antibacterial finish showed a large and clear inhibition zone on solid agar medium against E. coli and S. aureus, the width of the inhibition zone was 2.1 mm and 1.4 mm respectively. It indicated that the fabric after antibacterial finish was leaching type antibacterial fabric, and had good antibacterial effect. In the shake flask method, the finished fabric showed over 84.02% bacterial reduction against E. coli, and over 82.17% bacterial reduction against S. aureus. It is well recognized that lipopeptides has good antimicrobial activity.
The crude Lipopeptides as fabric softener were investigated. The bending length of wool fabric before softening finishes was 18.9 mm and after softening finishes was 13.7 mm. The results showed that lipopeptides as softener improved performance of wool soft.
引文
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