The Effect And Mechanism Of Alcohol Extract From Rhizome Gastrodia On Submerged Fermentation Of Grifola Frondosa

This arcticle aims at studying the effect and mechanism of alcohol extract from rhizome gastrodia on submerged fermentation of Grifola frondosa. The main contents in this paper includes the impact of extract from rhizome gastrodia on Grifola frondosa fermentation, transformation of the ingredients from gastrodia elata in the process of Grifola frondosa fermentation, the effect of p-hydroxybenzyl alcohol on the biosynthesis of exopolysaccharide produced by Grifola frondosa and its fermentation kinetics, and the preliminary study on glucose transferase in fermentation process.The qualitative and quantitative detection methods of gastrodine, p-hydroxybenzyl alcohol, p-hydroxybenzaldehyde and parishin were established. The ingredient variations of alcohol extract from gastrodia elata pre- and post- high pressure sterilization were analyzed. The conclusion demonstrated that the gastrodine content is 14.6955 mg/g, p-hydroxybenzyl alcohol content is 1.0857 mg/g, p-hydroxybenzaldehyde content is 1.9753 mg/g, parishin content is 2.0571 mg/g after high pressure sterilization.The influence on Grifola frondosa mycelial growth and extracellular polysaccharide of different concentrations of gastrodia elata alcohol extract showed as follows: with the addition of 7 %(v/v)(100mL fermented liquid contains 7mL gastrodia elata alcohol extract, namely 100 mL fermentation liquid contains around 0.7 g gastrodia elata), the amount of Grifola frondosa mycelium and EPS both reached maximum(3.35±0.09 g/L and 444.24±17.76 mg/L) respectively.The effects of each single component of the rhizoma gastrodiae alcohol extract on Grifola frondosa mycelial growth and extracellular polysaccharide were evaluated based on the amount of every single component in 7%(v/v) rhizoma gastrodiae alcohol extract. The additions consisted of Ⅰ gastrodia elata alcohol extract(7 g/L), Ⅱ gastrodin(103 mg/L), Ⅲ p-hydroxybenzyl alcohol(7.6 mg/L), Ⅳ p-hydroxybenzaldehyde(13.8 mg/L) and Ⅴ three kinds of single composition. The results showed that compared with the blank group, the five experiments could promote the growth of Grifola Frondosa and biosynthesis of extracellular polysaccharide to a certain extent. Ⅰand Ⅴ groups showed the same and maximal promoting effects. The Ⅱ group took the second place. But the Ⅲ and Ⅳ group demonstrated unconspicuous effects. The extracellular polysaccharide productivity showed that p-hydroxy benzaldehyde and p-hydroxybenzyl alcohol could improve the production ability of mycelium extracellular polysaccharide better compared with other groups.The amount variation of every component in rhizoma gastrodiae alcohol extract during the fermentation process of Grifola frondosa was analyzed. In 100 m L of the fermented liquid, the gastrodin decreased from 10.28 mg to 5.36 mg, reduced by 47.86%, p-hydroxybenzyl alcohol decreased from 0.76 to 0.67 mg, reduced by 11.84%, p-hydroxybenzaldehyde decreased from 1.38 to 0.35 mg, reduced by 74.64%, And only the parishin content increased, rose from 1.44 mg to 3.76 mg, increased by 161.11%. Resting cells transformation experiments were applied to further reveal that p-hydroxybenzyl alcohol and p-hydroxybenzaldehyde could be transformed into gastrodin in Grifola frondosa fermentation. The possible way was that p-hydroxybenzyl alcohol transformated in gastrodin by glycosylation in Grifola frondosa mycelium suspension, and p-hydroxybenzaldehyde was reduced in Grifola frondosa mycelium suspension, and then a small amount of gastrodin was synthesized.The influence of p-hydroxybenzyl alcohol on the synthesis of extracellular polysaccharide was studied. The results showed that the influence on Grifola frondosa mycelial growth and extracellular polysaccharide synthesis was obvious when p-hydroxybenzyl alcohol content was 200 mg/L. Compared with the blank group(without p-hydroxybenzyl alcohol), the mycelial increased by 22.73% and the EPS increased by 22.73%. In addition, the mycelium growth, reducing sugar content, extracellular polysaccharide content, pH value, the hydroxybenzene methanol, the changes of the content of gastrodin in submerged culture of Grifola frondosa and fermentation dynamics were studied.The enzymatic property of glucosyltransferase from Grifola frondosa was studied. The optimal reaction temperature was 50 ℃. Thermal stability of glucosyltransferase was decreased with the increasing of temperature. It was easily inactivated in high temperature. The optimum pH was 6. Glucosyltransferase was neither acid nor alkali resistance, It was also easily inactivated in partial acid and alkali environment.