Bafana Richa, Sivanesan Sarvanadevi, Pandey R A
AcSIR (Academy of Scientific and Innovative Research), CSIR-NEERI (National Environmental Engineering Research Institute), Nagpur, 440020 India.
Indian J Microbiol. 2017 Sep;57(3):322-328. doi: 10.1007/s12088-017-0661-5. Epub 2017 Jul 10.
Several fungi and starch-rich industrial residues were screened for itaconic acid (IA) production. Out of 15 strains, only three fungal strains were found to produce IA, which was confirmed by HPLC and GC-MS analysis. These strains were identified as strains C1 and C2, and strain C3 by sequencing of 18S rRNA gene and internal transcribed spacer regions. -aconitate decarboxylase () gene, which encodes a key enzyme in IA production in , was characterized from strains C1 and C2. C1 and C2 gene sequences showed about 96% similarity to the only available GenBank sequence of gene. 3-D structure and -aconitic acid binding pocket of Cad enzyme were predicted by structural modeling. Rice, corn and potato starch wastes were screened for IA production. These materials were enzymatically hydrolyzed under experimentally optimized conditions resulting in the highest glucose production of 230 mg/mL from 20% potato waste. On comparing the production potential of selected strains with different wastes, the best IA production was achieved with strain C1 (255.7 mg/L) using potato waste. Elemental composition as well as batch-to-batch variation in waste substrates were analyzed. The difference in IA production from two different batches of potato waste was found to inversely correlate with their phosphorus content, which indicated that produced IA under phosphate limiting condition. The potato waste hydrolysate was deionized to remove inhibitory ions like phosphate, resulting in improved IA production of 4.1 g/L by C1 strain, which is commercially competitive.
筛选了几种真菌和富含淀粉的工业废渣用于生产衣康酸(IA)。在15个菌株中,仅发现3个真菌菌株能够产生IA,通过高效液相色谱(HPLC)和气相色谱 - 质谱联用(GC - MS)分析得以证实。通过对18S rRNA基因和内部转录间隔区进行测序,将这些菌株鉴定为C1和C2菌株以及C3菌株。从C1和C2菌株中对编码IA生产关键酶的乌头酸脱羧酶()基因进行了表征。C1和C2的基因序列与GenBank中唯一可用的基因序列显示出约96%的相似性。通过结构建模预测了Cad酶的三维结构和乌头酸结合口袋。筛选了水稻、玉米和马铃薯淀粉废渣用于IA生产。在实验优化条件下对这些材料进行酶解,20%的马铃薯废渣产生的葡萄糖产量最高可达230 mg/mL。比较所选菌株利用不同废渣的生产潜力,使用马铃薯废渣时,C1菌株的IA产量最高(255.7 mg/L)。分析了废渣底物的元素组成以及批次间的差异。发现两批不同马铃薯废渣的IA产量差异与它们的磷含量呈负相关,这表明在磷酸盐限制条件下产生IA。对马铃薯废渣水解产物进行去离子处理以去除诸如磷酸盐等抑制性离子,C1菌株的IA产量提高到4.1 g/L,具有商业竞争力。