National Institute for Agro-environmental Sciences (NIAES), 3-1-3 Kannondai, Tsukuba, Ibaraki 305-8604, Japan; Research Fellow of the Japan Society for the Promotion of Science, Japan.
National Institute for Agro-environmental Sciences (NIAES), 3-1-3 Kannondai, Tsukuba, Ibaraki 305-8604, Japan.
J Biosci Bioeng. 2014 Mar;117(3):325-9. doi: 10.1016/j.jbiosc.2013.09.002. Epub 2013 Oct 3.
There is a need to speed up the degradation of used agricultural mulch films that are made of biodegradable plastics (BPs) in the field. Treating them with BP-degrading enzymes could be a solution to this problem. A cutinase-like enzyme of yeast Pseudozyma antarctica (PaE) has wide specificity of BPs degradation, but needs to be produced efficiently. Here we report that the production of PaE by P. antarctica can be increased by using xylose as carbon source. BP-degradation activity was analyzed using a polybutylene succinate-co-adipate (PBSA) emulsion as the substrate. Strain P. antarctica GB-4(1)W was found to be the best PaE producer among the tested strains. Using a 5-L jar fermentor with xylose fed-batch cultivation, high PaE productivity could be maintained and about 21 U/ml of PaE was obtained in 120 h. This amount was 100 times higher than the amount that we obtained previously (0.21 U/ml by flask cultivation using glycerol as carbon source). Under repeated xylose fed-batch cultivation with 24 h intervals, the maximum PaE production rate (0.34 U/ml/h) was maintained and the highest PaE productivity (28,000 U/2 L/d) was repeatedly obtained for 7 intervals. The activity of filtered jar-culture (crude PaE) was stable over 12 weeks at 4°C. Commercially available BP mulch films (20 μm thickness, cut into 1-cm-squares) were completely degraded by submerging them in crude PaE (2 U/ml) at 30°C in 24 h. These results indicated that concentrated PaE can rapidly degrade the strength of BP mulch films in the field so that they do not interfere with plowing.
需要加快田间可生物降解塑料(BP)农用覆盖膜的降解速度。用 BP 降解酶处理它们可能是解决这个问题的一种方法。来自南极假丝酵母(Pseudozyma antarctica)的一种角质酶样酶(PaE)对 BP 降解具有广泛的特异性,但需要高效生产。在这里,我们报告说,使用木糖作为碳源可以提高南极假丝酵母产生 PaE 的效率。使用聚丁二酸丁二醇酯-co-己二酸酯(PBSA)乳液作为底物来分析 BP 降解活性。结果发现,在所测试的菌株中,P. antarctica GB-4(1)W 是最好的 PaE 产生菌。使用带有木糖补料分批培养的 5-L 搅拌罐发酵罐,可以维持高 PaE 生产力,在 120 h 内获得约 21 U/ml 的 PaE。这一数量比我们之前获得的数量(使用甘油作为碳源的摇瓶培养获得的 0.21 U/ml)高 100 倍。在间隔 24 h 的重复木糖补料分批培养下,维持了最大的 PaE 生产速率(0.34 U/ml/h),并重复获得了 7 次最高的 PaE 生产力(28,000 U/2 L/d)。在 4°C 下,过滤罐培养(粗 PaE)的活性在 12 周内保持稳定。将商用 BP 覆盖膜(20 μm 厚,切成 1 cm 见方的小块)完全浸入 2 U/ml 的粗 PaE(2 U/ml)中,在 30°C 下 24 h 即可完全降解。这些结果表明,浓缩的 PaE 可以快速降解 BP 覆盖膜在田间的强度,使其不干扰耕作。