Highly efficient expression of human extracellular superoxide dismutase (rhEcSOD) with ultraviolet ‐B‐induced damage‐resistance activity in transgenic silkworm cocoons

The present study reports an efficient strategy for producing active rhEcSOD by transgenic silkworms. This strategy demonstrates that the rhEcSOD was successfully synthesized with high expression level and purified from cocoons with potent enzymatic activity against the ultraviolet-B-induced damage, thereby offering an effective alternative for producing the rhEcSOD with enzymatic and biological activities for the biomedical applications. AbstractExtracellular superoxide dismutase (EcSOD) protects tissues from oxidative stress, and thus is considered as a therapeutic agent for many diseases such as atherosclerosis, hypertension, and cancer. However, cost-effective production of bioactive recombinant human EcSOD (rhEcSOD) remains a challenge. Herein, we developed an efficient strategy for producing active rhEcSOD by transgenic silkworms. rhEcSOD was successfully synthesized as homodimers and homotetramers in the middle silk gland and spun into the cocoons with a concentration of 9.48  ± 0.21 mg/g. Purification of rhEcSOD from the cocoons could be conveniently achieved with a purity of 99.50% and a yield of 3.5 ± 0.5 mg/g. Additionally, N-glycosylation at the only site of N89 in rhEcSOD with 10 types were identified. The purified rhEcSOD gained the potent enzymatic activi ty of 4 162 ± 293 U/mg after Cu/Zn ions incorporation. More importantly, rhEcSOD was capable of penetrating and accumulating in the nuclei of cells to maintain cell morphology and attenuate ultrav...

Source: Insect Science - November 28, 2023 Category: Biology Authors: Feng Wang, Ri ‐Yuan Wang, De‐Bin Zhong, Ping Zhao, Qing‐You Xia Tags: ORIGINAL ARTICLE Source Type: research