Gedikli Serap, Güngör Gökhan, Toptaş Yağmur, Sezgin Dilber Ece, Demirbilek Murat, Yazıhan Nuray, Aytar Çelik Pınar, Denkbaş Emir Baki, Bütün Vural, Çabuk Ahmet
a Department of Biology, Graduate School of Natural and Applied Sciences , Eskisehir Osmangazi University , Eskisehir , Turkey.
b Department of Biotechnology and Biosafety, Graduate School of Natural and Applied Sciences , Eskisehir Osmangazi University , Eskisehir , Turkey.
Prep Biochem Biotechnol. 2018;48(7):610-618. doi: 10.1080/10826068.2018.1476885. Epub 2018 Jun 14.
In the present study, culture conditions of was optimized through Box-Behnken experimental design for hyaluronic acid production. About 0.87 gL of hyaluronic acid was produced under the determined conditions and optimal conditions were found as 38.42 °C, 24 hr and 250 rpm. The validity and practicability of this statistical optimization strategy were confirmed relation between predicted and experimental values. The hyaluronic acid obtained under optimal conditions was characterized. The effects of different conditions such as ultraviolet light, temperature and enzymatic degradation on hyaluronic acid produced under optimal conditions were determined. 118 °C for 32 min of autoclaved HA sample included 63.09 µg mL of d-glucuronic acid, which is about two-fold of enzymatic effect. Cytotoxicity of hyaluronic acid on human dermal cells (HUVEC, HaCaT), L929 and THP-1 cells was studied. effect on pro or anti-inflammatory cytokine release of THP-1 cells was determined. Although it varies depending on the concentration, cytotoxicity of hyaluronic acid is between 5 and 30%. However, it varies depending on the concentration of hyaluronic acid, TNF-α release was not much increased compared to control study. Consequently, purification procedure is necessary to develop and it is worth developing the bacterial hyaluronic acid.
在本研究中，通过Box-Behnken实验设计优化了透明质酸生产的培养条件。在确定的条件下产生了约0.87 g/L的透明质酸，最佳条件为38.42 °C、24小时和250转/分钟。通过预测值与实验值之间的关系证实了这种统计优化策略的有效性和实用性。对在最佳条件下获得的透明质酸进行了表征。确定了不同条件如紫外线、温度和酶降解对在最佳条件下产生的透明质酸的影响。在118 °C下高压灭菌32分钟的HA样品中含有63.09 μg/mL的d-葡萄糖醛酸，约为酶促作用的两倍。研究了透明质酸对人真皮细胞（HUVEC、HaCaT）、L929和THP-1细胞的细胞毒性。确定了其对THP-1细胞促炎或抗炎细胞因子释放的影响。尽管其因浓度而异，但透明质酸的细胞毒性在5%至30%之间。然而，其因透明质酸浓度而异，与对照研究相比，TNF-α的释放增加不多。因此，有必要开发纯化程序，并且开发细菌透明质酸是值得的。
