Enzymatic Modification of Starch Using Recombinant Genes from Sorghum in Escherichia coli: Insights and Potential Applications.

Abstract

SBEIIb (Sobic.004G163700), SSSIIa (Sobic.010G093400), and GBSSI (Sobic.010G022600) genes that regulate starch synthesis in sorghum endosperm were transferred into Escherichia coli by transgenic technology. SBEIIb, SSSIIa, and GBSSI enzymes were separated and purified through a Ni column and analyzed by electrophoresis with molecular weights and activities of 91.57 84.57, and 66.89 kDa and 551 and 700 and 587 U/μL, respectively. Furthermore, they were applied to starch modification, yielding interesting findings: the A chain content increased from 25.79 to 89.55% for SBEIIb-treated waxy starch, while SSSIIa extended the A chain to form DPs of the B chain, with A chain content decreasing from 89.55 to 37.01%, whereas GBSSI was explicitly involved in the synthesis of B1 chain, with its content increasing from 9.59 to 48.45%. Modified starch was obtained, which could be accurately applied in various industries. For instance, we prepared a sample (containing 89.6% A chain content) with excellent antiaging and antidigestion properties through SBEIIb modification. Moreover, higher RS3 (34.25%) and SDS contents (15.75%) of starch were obtained through the joint modification of SBEIIb and SSSIIa. These findings provide valuable insights for developing sorghum starch synthesis-related enzymes and offer opportunities for improving starch properties through enzymatic approaches.