Biotechnology, Bioprocess and Biocatalysis Group, Institute of Food Science and Technology, Federal University of Rio Grande do Sul, Av. Bento Gonçalves, 9500, P.O. Box 15090, ZC 91501-970, Porto Alegre, RS, Brazil; Department of Biocatalysis, ICP-CSIC, Campus UAM-CSIC, Cantoblanco, ZC 28049, Madrid, Spain.
Department of Biocatalysis, ICP-CSIC, Campus UAM-CSIC, Cantoblanco, ZC 28049, Madrid, Spain.
Enzyme Microb Technol. 2020 Jan;132:109397. doi: 10.1016/j.enzmictec.2019.109397. Epub 2019 Aug 9.
Pectin lyase (from Rohapect 10 L) was immobilized on glutaraldehyde supports at low ionic strength at pH 5, 6.5 or 8 and later incubated at pH 8 for 48 h. The activity recovery of the biocatalysts versus pectin was quite low, under 10% for all of the immobilized biocatalyst at 20 °C. However, a high stabilization was found when the enzyme was immobilized at pH 5, (e.g., the immobilized enzyme kept 83% of the activity when the free enzyme was fully inactivated (pH 4.8 and 55 °C in 5 h)). This biocatalyst increased the activity versus pectin in an almost exponential way when temperature increased until reach the maximum temperature used in the study (90 °C), conditions where the free enzyme was almost inactive. The immobilized biocatalyst was also active even at pH 9, where the free enzyme was fully inactive. This biocatalyst could be reused for pectin hydrolysis 5 times for 72 h reaction cycles at 40 °C maintaining more than 90% of the initial activity.
果胶裂解酶（来自 Rohapect 10L）在低离子强度和 pH5、6.5 或 8 下固定在戊二醛载体上，随后在 pH8 下孵育 48 小时。所有固定化生物催化剂在 20°C 下对果胶的活性回收率都相当低，低于 10%。然而，当酶在 pH5 下固定时，发现酶有很高的稳定性（例如，当游离酶完全失活（pH4.8 和 55°C 下 5 小时）时，固定化酶保持 83%的活性）。这种生物催化剂在升高温度时几乎呈指数级增加对果胶的活性，直到达到研究中使用的最高温度（90°C），在这种条件下，游离酶几乎没有活性。固定化生物催化剂在 pH9 时也具有活性，而游离酶在 pH9 时完全失活。该固定化生物催化剂在 40°C 下进行 72 小时的反应循环，可重复用于果胶水解 5 次，保持初始活性的 90%以上。
