Microbial fermentation of botanicals

Abstract

A method for microbial fermentation of botanicals includes steps of: fermenting with wall-breaking fungi, and then fermenting with probiotics.

Claims

1-10. (canceled)

11: A microbial fermentation method of botanicals, comprising steps of: fermenting with wall-breaking fungi, and then fermenting with probiotics.

12: The microbial fermentation method, as recited in claim 11, wherein the wall-breaking fungi are wood-grown fungi and/or Cordyceps.

13: The microbial fermentation method, as recited in claim 11, wherein the wall-breaking fungi are selected from a group consisting of Ganoderma, Poria cocas, Grifola umbellata, Cordyceps sinensis and/or Cordyceps militaris.

14: The microbial fermentation method, as recited in claim 12, wherein the wall-breaking fungi are selected from a group consisting of Ganoderma, Poria cocos, Grifola umbellata, Cordyceps sinensis and/or Cordyceps militaris.

15: The microbial fermentation method, as recited in claim 11, wherein the probiotics are selected from a group consisting of Saccharomyces, Pichia, Hansenula, Lactobacillus, Bifidobacterium, Leuconostoc, Streptococcus, Lactocaccusand Acetobacterium.

16: The microbial fermentation method, as recited in claim 14, wherein the probiotics are selected from a group consisting of Saccharomyces, Pichia, Hansenula, Lactobacillus, Bifidobacterium, Leuconostoc, Streptococcus, Lactococcus and Acetobacterium.

17: The microbial fermentation method, as recited in claim 15, wherein the probiotics are selected from a group consisting of Saccharomyces cerevisiae, Saccharomyes eerevisiae, Pichia pastoris, Pichia ohmeri, Pichia membranaefaciens, Hansenula anomala, Lactobacillus buchneri, Lactobacillus panis, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus fermenti, Lactobacillus salivarius, Lactobacillus breves, Lactobacillus veridescens, Lactobacillus delbrueckii bulgaricus, Lactobacillus rhamnosus, Lactobacillus cellobiosus, Lactobacillus casei subsp. casei, Lactobacillus helveticus subsp. jugurti, Lacticum Jansen, Lactobacillus Pentosus, Acetobacter pasteuranus, Acetobacter rancens, Bifidobacterium adolescents, Bifidobacterium bifidum, Bifidobacterium infantis, Bifidobacterium longum, Leuconostoc mesenteroides aureus, Streptococcus salivarius subsp. thermophilus and Lactococcus lactis subsp. Cremoris.

18: The microbial fermentation method, as recited in claim 16, wherein the probiotics are selected from a group consisting of Saccharomyces cerevisiae, Saccharomyes eerevisiae, Pichia pastoris, Pichia ohmeri, Pichia membranaefaciens, Hansenula anomala, Lactobacillus buchneri, Lactobacillus panis, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus fermenti, Lactobacillus salivarius, Lactobacillus breves, Lactobacillus veridescens, Lactobacillus delbrueckii bulgaricus, Lactobacillus rhamnosus, Lactobacillus cellobiosus, Lactobacillus casei subsp. casei, Lactobacillus helveticus subsp. jugurti, Lacticum Jansen, Lactobacillus Pentosus, Acetobacter pasteuranus, Acetobacter rancens, Bifidobacterium adolescents, Bifidobacterium bifidum, Bifidobacterium infantis, Bifidobacterium longum, Leuconostoc mesenteroides aureus, Streptococcus salivarius subsp. thermophilus and Lactococcus lactis subsp. Cremoris.

19: The microbial fermentation method, as recited in claim 17, wherein the probiotics comprise yeast, Lactobacillus and Acetobacterium.

20: The microbial fermentation method, as recited in claim 18, wherein the probiotics comprise yeast, Lactobacillus and Acetobacterium.

21. The microbial fermentation method, as recited in claim 19, wherein mixed fermentation of yeast and Lactobacillus is followed by fermentation of Acetobacterium.

22: The microbial fermentation method, as recited in claim 20, wherein mixed fermentation of yeast and Lactobacillus is followed by fermentation of Acetobacterium.

23: The microbial fermentation method, as recited in claim 11, wherein the botanical is ginseng.

24: The microbial fermentation method, as recited in claim 22, wherein the botanical is ginseng.

25: The microbial fermentation method, as recited in claim 11, wherein the botanical is pueraria.

26: The microbial fermentation method, as recited in claim 22, wherein the botanical is pueraria.

27: A product prepared by a microbial fermentation method of botanicals as recited in claim 11.

28: A product prepared by a microbial fermentation method of botanicals as recited in claim 26.

Description

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

[0039] The following embodiments are intended to describe the present invention in detail, and are not intended to limit the scope of the present invention. Modifications and adjustments made by those skilled in the art based on the disclosure also belong to the scope of the present invention.

Embodiment 1

[0040] Weighing 10 Kg of ginseng, pulverizing to 30 meshes, pouring into a fermenting tank, adding 80 Kg of culture solution (by mass ratio: glucose 2.4%, yeast extract 0.2%, peptone 0.4%, potassium dihydrogen phosphate 0:1%, magnesium sulfate 0.1%, and distilled water 96.8%), thoroughly mixing, adjusting pH to 6.8 with sodium hydroxide solution, sterilizing with steam for 30 min; cooling to room temperature, and inoculating Ganoderma lucidum (Xiangchizhi No. 1, Hunan Province non-major crop variety registration certificate number XPD010-2013, obtained from State Key Laboratory of Sub-health Intervention Technology), keeping a stirring speed in the fermentation tank at 120 rpm, keeping the temperature at 20 C., and fermenting for 15 days; then adding 1% (total raw material mass fraction) yeast (Pichia ohmeri, CGMCC No. 2.1803, millions of viable bacteria) and 1% (total raw material mass fraction) Lactobacillus (Lactobacillus plantarum, CGMCC No. 1.6971, millions of viable bacteria), and statically fermenting at 28 C. for 7 days; then adding 1% (total raw material mass fraction) Acetobacterium (Acetobacter rancens AS.1.41, purchased from Shanghai No. 1 Brewing Plant, millions of viable bacteria), statically fermenting at 30 C. for 7 days; filtering and collecting liquid.

[0041] According to test, the ginseng liquid obtained by the present invention has a total ginsenoside content of 1.06 mg/mL (the determination method refers to that in the paper of Content determination of total ginsenoside, Gag Liping et al., Journal of Zhejiang Institute of Engineering, 2012, 31(3): 382-388), which is 7.01 times higher than the conventional decoction (a weight ratio of ginseng and water is 1:8). Furthermore, 0.75 mg/mL Ganoderma triterpenoid content is also detected in the liquid (the assay method refers to that in Chinese Pharmacopoeia 2015 edition). All data are average values of triplicate samples.

Embodiment 2

[0042] Weighing 10 Kg of ginseng, pulverizing to 30 meshes, pouring into a fermenting tank, adding 80 Kg, of the culture solution as mentioned in the embodiment 1, thoroughly mixing, adjusting pH to 6.8 with sodium hydroxide solution, sterilizing with steam for 30 min; cooling to room temperature, and inoculating Ganoderma (Ganoderma valesiacum, CGMCC No. 5.772, belonging to Ganoderma sinense), keeping a stirring speed in the fermentation tank at 100 rpm, keeping temperature at 23 C., and fermenting for 15 days; then adding 1% (total raw material mass fraction) yeast (Saccharomyces cerevisiae, CGMCC No. 2.3888, millions of viable bacteria) and 1% (total raw material mass fraction) Lactobacillus (Lactobacillus panis, CGMCC No. 1.3925, millions of viable bacteria), and statically fermenting at 28 C. for 7 days; then adding 1% (total raw material mass fraction) Acetobacterium (Acetobacter rancens AS.1.41, millions of viable bacteria), statically fermenting at 30 C. for 7 days; filtering and collecting liquid.

[0043] According to test, the ginseng liquid obtained by the present invention has a total ginsenoside content of 1.03 mg/mL (the determination method as mentioned in the embodiment 1), which is 6.81 times higher than the conventional decoction (a weight ratio of ginseng and water is 1:8). Furthermore, 0.64 mg/mL Ganoderma triterpenoid content is also detected in the liquid (the assay method as mentioned in the embodiment 1). All data are average values of triplicate samples.

Embodiment 3

[0044] Weighing 10 Kg of pueraria, pulverizing to 20 meshes, pouring into a fermenting tank, adding 100 Kg of culture solution as mentioned in the embodiment 1, thoroughly mixing, adjusting pH to 6.5 with sodium hydroxide solution, sterilizing at 121 C. for 30 min; cooling to room temperature, and inoculating Cordyceps (Xiangbeichongcao No. 1, Hunan Province non-major crop variety registration certificate number XPD009-2013, obtained from State Key Laboratory of Sub-health Intervention Technology), keeping a stirring speed in the fermentation tank at 150 rpm, keeping temperature at 25 C., and fermenting for 10 days; then adding 3% (total raw material mass fraction) yeast (Saccharomyces cerevisiae, CGMCC No. 2.3973, millions of viable bacteria) and 1% (total raw material mass fraction) Lactobacillus (Lactobacillus buchneri, CGMCC No. 1.3114, millions of viable bacteria), and statically fermenting at 28 C. for 8 days; then adding 1% (total raw material mass fraction) Acetobacterium (Acetobacter pasteuranus 1.01, millions of viable bacteria), statically fermenting at 30 C. for 7 days; filtering and collecting liquid.

[0045] According to test, the pueraria liquid obtained by the present invention has a puerarin content of 0.119 mg/mL (the determination method refers to that in Health Foods of Chinese National Standard GB/T22251-2008), which is 1.25 times higher than the conventional decoction (a weight ratio of pueraria and water is 1:10). Furthermore, 11.3 g/mL cordycepin content is also detected in the liquid (the assay method refers to that in Screening of high-yield strains of cordycepin and effects of different additives on the yield of cordycepin, Wang Lei et al., Journal of Fungal Materials, 2012, 31(3): 382-388). All data are average values of triplicate samples.

Embodiment 4

[0046] Weighing 10 Kg of pueraria, pulverizing to 20 meshes, pouring into a fermenting tank, adding 100 Kg of the culture solution as mentioned in the embodiment 1, thoroughly mixing, adjusting pH to 6.5 with sodium hydroxide solution, sterilizing at 121 C. for 30 min; cooling to room temperature, and inoculating Cordyceps (Cordyceps militaris, CGMCC No. 5.856), keeping a stirring speed in the fermentation tank at 130 rpm, keeping temperature at 28 C., and fermenting for 10 days; then adding 3% (total raw material mass fraction) yeast (Pichia membranaefaciens, CGMCC No. 2.661, millions of viable bacteria) and 1% (total raw material mass fraction) Lactobacillus (Lactobacillus panis, CGMCC No. 1.3925, millions of viable bacteria), and statically fermenting at 28 C. for 8 days; then adding 1% (total raw material mass fraction) Acetobacterium (Acetobacterium Huniang 1.01, millions of viable bacteria), statically fermenting at 30 C. for 7 days; filtering and collecting liquid.

[0047] According to test, the pueraria. liquid obtained by the present invention has a puerarin content of 0.113 mg/mL (the determination method as mentioned in the embodiment 3), which is 1.19 times higher than the conventional decoction (a weight ratio of pueraria and water is 1:10). Furthermore, 13.7 g/mL cordycepin content is also detected in the liquid (the assay method as mentioned in the embodiment 3). All data are average values of triplicate samples.