Hwang Hee Sun, Lee Sung Hak, Baek Yu Mi, Kim Sang Woo, Jeong Yong Kee, Yun Jong Won
Department of Biotechnology, Daegu University, Kyungsan, Kyungbuk, 712-714, South Korea.
Appl Microbiol Biotechnol. 2008 Mar;78(3):419-29. doi: 10.1007/s00253-007-1329-6. Epub 2008 Jan 10.
In the present study, optimum culture conditions for the production of extracellular polysaccharides (EPS) in submerged culture of an edible mushroom, Laetiporus sulphureus var. miniatus and their stimulatory effects on insulinoma cell (RINm5F) proliferation and insulin secretion were investigated. The maximum mycelial growth (4.1 g l(-1)) and EPS production (0.6 g l(-1)) in submerged flask culture were achieved in a medium containing 30 g l(-1) maltose, 2 g l(-1) soy peptone, and 2 mM MnSO(4).5H2O at an initial pH 2.0 and temperature 25 degrees C. In the stirred-tank fermenter under optimized medium, the concentrations of mycelial biomass and EPS reached a maximum level of 8.1 and 3.9 g l(-1), respectively. Interestingly, supplementation of deep sea water (DSW) into the culture medium significantly increased both mycelial biomass and EPS production by 4- and 6.7-fold at 70% (v/v) DSW medium, respectively. The EPS were proved to be glucose-rich polysaccharides and were able to increase proliferation and insulin secretary function of rat insulinoma RINm5F cells, in a dose-dependent manner. In addition, EPS also strikingly reduced the streptozotocin-induced apoptosis in RINm5F cells indicating the mode of the cytoprotective role of EPS on RINm5F cells.
在本研究中,对食用真菌硫黄多孔菌微小变种液体培养生产胞外多糖(EPS)的最佳培养条件及其对胰岛素瘤细胞(RINm5F)增殖和胰岛素分泌的刺激作用进行了研究。在初始pH值为2.0、温度为25℃的含有30 g l(-1)麦芽糖、2 g l(-1)大豆蛋白胨和2 mM MnSO₄·5H₂O的培养基中,摇瓶液体培养获得了最大菌丝体生长量(4.1 g l(-1))和EPS产量(0.6 g l(-1))。在优化培养基的搅拌罐发酵器中,菌丝体生物量和EPS浓度分别达到最大水平8.1和3.9 g l(-1)。有趣的是,在培养基中添加深海海水(DSW),当DSW培养基体积分数为70%(v/v)时,菌丝体生物量和EPS产量分别显著增加了4倍和6.7倍。已证明EPS是富含葡萄糖的多糖,能够以剂量依赖的方式增加大鼠胰岛素瘤RINm5F细胞的增殖和胰岛素分泌功能。此外,EPS还显著降低了链脲佐菌素诱导的RINm5F细胞凋亡,表明EPS对RINm5F细胞具有细胞保护作用。