Hwang H J, Kim S W, Xu C P, Choi J W, Yun J W
Department of Biotechnology, Daegu University, Kyungsan, Kyungbuk, Korea.
J Appl Microbiol. 2003;94(4):708-19. doi: 10.1046/j.1365-2672.2003.01903.x.
The objective of the present study was to determine the optimal culture conditions for the production of four groups of exopolysaccharides (EPSs) in Phellinus gilvus by submerged culture and to investigate their molecular properties by multi-angle laser-light scattering (MALLS) analysis.
The optimal temperature and initial pH for the production of both mycelial biomass and EPSs in P. gilvus by submerged flask cultures were found to be 30 degrees C and pH 9.0, respectively. Glucose and corn steep powder were the most suitable carbon and nitrogen source for both mycelial biomass and EPS production. Optimal medium composition was determined to be glucose 30 g l-1, corn steep powder 5 g l-1, MgSO4 1.23 g l-1, KH2PO4 0.68 g l-1, and K2HPO4 0.87 g l-1. Four groups of EPSs (Fr-I, II, III, and IV) were obtained from the culture filtrates by gel filtration chromatography on Sepharose CL-4B and characterized by size exclusion chromatography (SEC) coupled with MALLS. The weight average molar mass (Mw) of Fr-I, Fr-II, Fr-III and Fr-IV were determined to be 8.628 x 106 (+/-129 420), 1.045 x 106 (+/-19 855), 61.09 x 104 (+/-1244), and 33.55 x 104 (+/-134) g mol-1, respectively.
Under optimal culture conditions, the maximum EPS production in a 5-l stirred fermenter indicated 5.3 g l-1 after 11 days of fermentation. The SEC/MALLS analysis revealed that Fr-I, which has extremely high molecular weight, was presumably an aggregate of complex polysaccharides forming a compact globular shape; whereas Fr-II was nearly spherical, Fr-III and Fr-IV were rod-like chains in an aqueous solution.
This is the first report on the production of high amounts of EPSs from liquid-culture of the basidiomycete, P. gilvus. The SEC/MALLS approach used in this study could be useful in providing greater insight into the characterization of the mushroom polysaccharides without carrying out elaborate fractionation procedures prior to analysis.
本研究的目的是通过深层培养确定桑黄中四组胞外多糖(EPSs)产生的最佳培养条件,并通过多角度激光散射(MALLS)分析研究其分子特性。
通过摇瓶深层培养发现,桑黄中菌丝体生物量和EPSs产生的最佳温度和初始pH分别为30℃和pH 9.0。葡萄糖和玉米浆是菌丝体生物量和EPS产生最合适的碳源和氮源。确定最佳培养基组成为葡萄糖30 g l-1、玉米浆5 g l-1、MgSO4 1.23 g l-1、KH2PO4 0.68 g l-1和K2HPO4 0.87 g l-1。通过Sepharose CL-4B凝胶过滤色谱从培养滤液中获得四组EPSs(Fr-I、II、III和IV),并通过尺寸排阻色谱(SEC)结合MALLS进行表征。Fr-I、Fr-II、Fr-III和Fr-IV的重均摩尔质量(Mw)分别确定为8.628×106(±129420)、1.045×106(±19855)、61.09×104(±1244)和33.55×104(±134)g mol-1。
在最佳培养条件下,5升搅拌发酵罐中发酵11天后EPS的最大产量为5.3 g l-1。SEC/MALLS分析表明,分子量极高的Fr-I可能是形成紧密球状的复合多糖聚集体;而Fr-II接近球形,Fr-III和Fr-IV在水溶液中为棒状链。
这是关于担子菌桑黄液体培养产生大量EPSs的首次报道。本研究中使用的SEC/MALLS方法有助于在不进行复杂分级分离程序的情况下,更深入地了解蘑菇多糖的特性。