【作者】 李永泉； 谢志鹏； 潘海峰； 叶兴乾； 张建国；

【Author】 Yongquan Li;Zhipeng Xie;Haifeng Pan;Xingqian Ye;Jianguo Zhang;Institute of Biochemistry,Zhejiang University;School of biosystems engineering and food science;

【机构】 浙江大学生物化学研究所； 浙江大学生工食品学院；

【摘要】 L-酒石酸是有机酸产业领域的重要产品,主要用作葡萄酒与饮料的酸味剂、食品乳化剂的合成原料(DATEM)等,但目前L-酒石酸批式生物合成技术存在的酶活力低、酶稳定性差、催化效率不高、固定化细胞传质效率低、工艺流程长、污染重及底物与产物的抑制效应等问题。本项目采用基因重组技术重构了L-酒石酸合成酶,在解决了固定化细胞通透性、表面结垢等关键问题基础上,构建了高催化性能的含酶细胞载体,产物对底物的摩尔转化率高达95%,固定化酶可重复使用10次以上;通过反应与分离耦合,降低了底物与产物的抑制效应,酶促反应效率由0.5kg提高到0.7kg产物/kg细胞/小时,吨产品原料成本降低38.8%、污染排放(COD)降低32.2%;。最终成功研制了L-酒石酸高效生物合成技术,建立了连续化生产线,实现了产业化,2013年产业化实施企业宝晶股份有限公司生产的L-酒石酸销售额逾3亿,全球市场占有率为20%以上。当我国有机酸产品普遍遭遇国际反倾销诉讼时,公司依托创新技术于2006年和2012年两次赢得欧盟对华酒石酸的反倾销诉讼,成为获"零反倾销税率"待遇的国内唯一企业,是目前国际最大的酒石酸生产商。本项目发表论文15篇,申请和授权发明专利7项,评为"国家重点新产品"称号,获浙江省科学技术奖一等奖和教育部科技进步奖一等奖。更多还原

【Abstract】 L-tartaric acid,an important product in organic acid industry,is mainly used as acidulants in wine and beverage,synthetic materials of food emulsifier(DATEM) and etc.However,batch biosynthesis of L-tartaric acid presently exist some disadvantages,such as low enzyme activity,poor stability of enzyme,unsatisfied catalytic efficiency,bad mass transfer efficiency of immobilized cells,long process flow,severe pollution,substrate-product inhibition and etc.In this project,L-tartaric acid synthetase was reconstructed by gene recombination and high catalytic enzyme-containing cell carriers were constructed,after resolving the key problems of immobilized cells permeability and fouling on the surface.The mole substrate conversion rate could reach up to 95%,and the immobilized enzyme could be reused more than 10 repeated batches.Through reaction-separation integrated control,substrate-product inhibition was cut down,the enzymatic reaction efficiency was increased from 0.5 to 0.7 kg product/kg cells/h,the raw material cost of product per ton was reduced 38.8%and the pollutant emission(COD) was decreased 32.2%.Finally,we successfully developed efficient biosynthesis of L-tartaric acid,constructed the continuous production line and realized industrialization.Hangzhou Bioking Biochemical Engineering Co.,Ltd was the enterprise to realize industrialization.In 2013,the sales volume of L-tartaric acid was about 0.3 billion and the global market share was more than 20%..When the organic acid producers in our country were generally suffered the international anti-dumping cases,Hangzhou Bioking Biochemical Engineering Co.,Ltd won the EU’s anti-dumping litigation of L-tartaric acid against China twice in 2006 and 2012,due to the technological innovation.It’s the only domestic enterprise with ’zero anti-dumping tax rate’ treatment and become the largest international manufacturers of tartaric acid.This project published 15 papers,applied and granted 7 invention patents,acquired the ’ national important new product’ title,won the first prize of Zhejiang province scientific and technological award and the first prize of ministry of education scientific and technological progress award.更多还原