NL2030836B1 - Ganoderma lucidum fermentative liquid food and preparation method thereof - Google Patents

Abstract

The present disclosure discloses a Ganoderma lucidum fermentative liquid food, and belongs to the field of foods. The Ganoderma lucidum fermentative liquid food includes a Ganoderma lucidum fermentative liquid and a Moringa seed liquid at a ratio of the Ganoderma lucidum fermentative liquid to the Moringa seed liquid of 1:1—5. A compound beverage of the Moringa seed liquid and the Ganoderma lucidum fermentative liquid prepared in the present disclosure is creamy white with a faint fragrance of Moringa seeds, the beverage is homogeneous and tastes sweet and sour.

Description

GANODERMA LUCIDUM FERMENTATIVE LIQUID FOOD AND

PREPARATION METHOD THEREOF

BACKGROUND ART

[01] Ganoderma lucidum is a precious edible and medicinal fungus with a long history and multiple physiological regulatory functions!!! As a fruiting body of fungus

Ganoderma lucidum belonging to the Polyporaceae family, Ganoderma lucidum has a high medicinal value, and can be used to improve immune function, inhibit tumor growth, soothe the liver and detoxify, lower blood pressure and blood lipid, and treat neurasthenia, palpitation and shortness of breath!?!.

[02] Fermentation conditions of Ganoderma lucidum, such as pH, temperature, inoculum dose, culture time and other culture conditions during fermentation are crucial for liquid fermentation of Ganoderma lucidum. Mycelia of Ganoderma lucidum prefer slightly acidic culture conditions, and can grow at a pH of 4.5-8.0"! and an optimum pH for mycelial growth is 5-6!" The mycelia of Ganoderma lucidum can grow at a temperature of 22-35 °C, and an optimum temperature for mycelial growth is 27- 30 °CH Yang Yan et al.b! studied effect of inoculum dose on mycelial biomass, and determined that an optimum inoculum dose was 10%. It is generally believed that an optimum age of mycelia is 4-5 d, and the optimal culture time should be 5 d.

[03] With the development of food science and the improvement of people’s living standard, people are paying more and more attention to nutritional and healthcare functions of foods. To improve the use value of Ganoderma lucidum, more researches on Ganoderma lucidum foods can be carried out.

SUMMARY

[04] In this experiment, a compound solution of a Ganoderma lucidum fermentative liquid and Moringa seeds was developed, which helps enrich varieties of Ganoderma lucidum foods and expand the market for Ganoderma lucidum foods.

[05] A Ganoderma lucidum fermentative liquid food, including a Ganoderma lucidum fermentative liquid and a Moringa seed liquid.

[06] Preferably, a ratio of the Ganoderma lucidum fermentative liquid to the Moringa seed liquid is 1:1-5.

[07] Preferably, the Ganoderma lucidum fermentative liquid food further includes an acidulant, a sweetener and a stabilizer.

[08] Preferably, the acidulant is malic acid.

[09] Preferably, the sweetener is xylitol.

[10] Preferably, the stabilizer is Arabic gum.

BRIEF DESCRIPTION OF THE DRAWINGS

[11] In order to explain the technical solutions in the embodiments of the present disclosure or in the prior art more clearly, the drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings in the following description are merely some embodiments of the present disclosure, and are intended to illustrate advantages of the technical solutions of the present disclosure.

[12] FIG. 1 is an image of slant culture of Ganoderma lucidum;

[13] FIG. 2 is an image of liquid culture of Ganoderma lucidum;

[14] FIG. 3 is a microscopic image of mycelia during fermentation of Ganoderma lucidum in liquid culture;

[15] FIG. 41s a standard curve of rutin content;

[16] FIG. 5 is a standard curve of gallic acid;

[17] FIG. 6 shows sensory evaluation scores of effect of acidulants on the compound beverage;

[18] FIG. 7 shows sensory evaluation scores of effect of sweeteners on the compound beverage; and

[19] FIG. 8 shows sensory evaluation scores of effect of stabilizers on the compound beverage.

DETAILED DESCRIPTION OF THE EMBODIMENTS Embodiment

Experimental materials

Materials and reagents

[20] Ganoderma lucidum strain, peptone, malt extract powder, agar, Moringa seeds, anhydrous citric acid, lactic acid, malic acid, tartaric acid, L-ascorbic acid (VC), sucralose, xylitol, mogroside, Arabic gum, guar gum, locust bean gum and konjac gum.

Experimental instruments

[21] Balance, autoclave, incubator, shaker, clean bench, microscope and UV-Vis spectrophotometer.

Experimental method

[22] 1. Preparation of culture medium

[23] Slant medium: 3 g of peptone, 20 g of malt extract powder, 12 g of agar and 600 mL of water are heated, melted and mixed well, then filled and autoclaved for 15 min.

[24] Liquid medium: 5 g of peptone, 30 g of malt extract powder and 1000 mL of water are filled in 500 mL conical flasks and autoclaved for 15 min.

[25] 2. Activation of Ganoderma lucidum strain

[26] Under sterile operation, a preserved Ganoderma lucidum strain is inoculated to a sterilized slant medium to fully activate the strain.

[27] 3. Liquid culture of Ganoderma lucidum

[28] Under sterile operation, activated Ganoderma lucidum mycelia are picked up and inoculated into a pre-sterilized liquid medium for liquid shake flask culture.

[29] 4. Microscopic observation of Ganoderma lucidum mycelia

[30] Ganoderma lucidum mycelial pellets cultured in a liquid medium are observed under a microscope. Photos are taken every day to record different forms of Ganoderma lucidum mycelia.

[31] 5. Determination of content of flavonoids in Ganoderma lucidum

[32] With rutin as a reference standard, content of flavonoids in Ganoderma lucidum is determined by NaNO2-AI(NO3)3-NaOH colorimetry with reference to Yang

Guangming et al. [6].

[33] 6. Determination of content of polyphenols in Ganoderma lucidum

[34] Content of polyphenols in Ganoderma lucidum is determined by Folin-Ciocalteu colorimetry with reference to Yang Xuanping et al. [7].

[35] 7. Culture in food-grade liquid medium

[36] Mycelial pellets that grow vigorously in a liquid medium are inoculated into a food-grade medium, and morphology is observed every day.

[37] 8. Production of compound beverage of Moringa seeds and Ganoderma lucidum

[38] 8.1 Preparation of Moringa seed liquid and compounding with Ganoderma lucidum fermentative liquid

[39] Moringa seeds are peeled and ground with 200, 250, 300, 350, 400, 450 and 500 mL of water to determine an optimum amount of water to be added. The Moringa seed liquid and the Ganoderma lucidum fermentative liquid are compounded at a ratio of 1:1,

1:2, 1:3, 1:4 and 1:5 to determine an optimum compound ratio.

[40] 8.2 Single factor experiment

[41] Optimum types and addition amount of acidulants, sweeteners and stabilizers in the compound solution are determined separately by sensory evaluation.

[42] 8.2.1 Acidulants

[43] L-ascorbic acid and anhydrous citric acid are added at 0.02%, 0.03%, 0.04%, 0.05% and 0.06%, and malic acid, tartaric acid and lactic acid are added at 0.01%, 0.02%, 0.03%, 0.04% and 0.05%.

[44] 8.2.2 Sweeteners

[45] Based on the above results, sucralose is added at 2%, 2.5%, 3%, 3.5% and 4%, and xylitol and mogroside are added at 2%, 3%, 4%, 5% and 6%, respectively.

[46] 8.2.3 Stabilizers

[47] Based on the above results, Arabic gum and guar gum are added at 0.2%, 0.3%, 0.4%, 0.5% and 0.6%, and locust bean gum and konjac gum are added at 0.2%, 0.25%, 0.3%, 0.35% and 0.4%, respectively.

[48] 8.2.4 Orthogonal experiment

[49] An orthogonal experimental table is designed with five levels for each factor to determine an optimum addition amount.

[50] 8.3 Sensory evaluation [S51] Beverages with different proportions are evaluated by sensory evaluation such as smell, sight and taste.

[52] Results and analysis

[53] 1. Image of slant culture of Ganoderma lucidum

[54] Ganoderma lucidum mycelia cultured on a slant are white fluffy, growing radially in all directions and eventually covering the whole slant, as shown in FIG. 1.

[55] 2. Image of Ganoderma lucidum cultured in a liquid

[56] Ganoderma lucidum mycelial pellets cultured in a liquid are milky white and are surrounded by a transparent circle, that is, exopolysaccharides,

[57] as shown in FIG. 2.

[58] 3. Image of Ganoderma lucidum mycelia observed under a microscope

[59] As the culture time goes by, branches grow gradually from Ganoderma lucidum mycelia with septa, and contents and spores can be clearly observed, as shown in FIG. 3.

[60] 4. Measured result of content of flavonoids

[61] With rutin as a reference standard, content of flavonoids in the Ganoderma lucidum fermentative liquid is determined. As shown in FIG. 4, rutin has a standard curve of y=0.0004x+0.0877. 5S Table 4-1 Content of flavonoids in samples

No. Samplesize/mlL Absorbance Content of flavonoids oor onze 2 1 0.121 0.269 3 1 0.143 0.442 4 1 0.151 0.508 1 0.155 0.547

[62] As shown in Table 4-1, as the culture time goes by, the content of flavonoids in

Ganoderma lucidum increases gradually at a slowing-down rate. The content is determined by spectrophotometry, which is characterized by simple operation, accurate result and high sensitivity.

[63] 5 Measured result of content of polyphenols

[64] With gallic acid as a reference standard, content of polyphenols in the

Ganoderma lucidum fermentative liquid 1s determined. As shown in FIG. 5, a standard curve is y=107.89x+0.0157.

Table 4-2 Content of polyphenols in samples

No. Samplesizeml Absorbance Comtent of polyphenols/mg/g 2 1 0.0854 1.76 3 1 0.1286 2.24 4 1 0.1684 2.48 5 1 0.2756 2.88 165] As shown in Table 4-2, mass concentrations of phenolic compounds show a good linear relationship with absorbance, and the content of polyphenols in Ganoderma lucidum gradually increases as the culture time goes by. The Folin-Ciocalteu colorimetry used has the advantages of simple operation and high accuracy.

[66] 6. Results of single factor experiment on compounded Moringa seeds and

Ganoderma lucidum fermentative liquid

[67] 6.1 Results of single factor experiment on acidulants

[68] According to sensory evaluation, an optimum water addition of the Moringa seed liquid is 400 mL, and an optimum compound ratio of the Moringa seed liquid to the fermentative liquid is 1:3. An optimum compound liquid is used for sensory evaluation of compound beverages with different acidulants and different additions. As shown in FIGS. 4-6, lactic acid and anhydrous citric acid will cause excessive acidity of a Moringa seed compound beverage, so that addition of lactic acid and anhydrous citric acid has some effect on a taste of the beverage and makes the Moringa seed compound beverage more astringent. A compound beverage added with tartaric acid and

L-ascorbic acid is not as tasty as that added with malic acid. Malic acid has a regulatory effect on the taste of the compound beverage, and a larger amount or smaller amount of malic acid added will affect the taste of the compound beverage. When the amount of malic acid added is small, the compound beverage is slightly sour with strong astringency. When the amount of malic acid added is large, the compound beverage is excessively sour with a discordant taste. As shown in FIG. 6, malic acid with an addition of 0.03% is the optimum.

[69] 6.2 Results of single factor experiment on sweeteners

[70] According to sensory evaluation on compound beverages with different sweeteners and different additions, as shown in FIG. 7, the taste of a compound beverage added with mogroside is light and not ideal with strong astringency. The taste of a compound beverage added with sucralose is slightly light, and cost is high. Xylitol has a regulatory effect on the taste of the compound beverage. When the amount of xylitol added is small, the compound beverage is slightly sweet with a faint and poor taste.

When the amount of xylitol added is large, the compound beverage is excessively sweet with a discordant taste. Therefore, according to sensory evaluation scores in FIG. 7, xylitol with an addition of 3% is the optimum.

[71] 6.3 Results of single factor experiment on stabilizers

[72] According to sensory evaluation on compound beverages with different stabilizers and different additions, as shown in FIGS. 4-8, a compound beverage added with locust bean gum is less stable than that added with Arabic gum. Arabic gum has a regulatory effect on stability of the compound beverage. When the amount of Arabic gum added is small, the compound beverage is unstable, so that the beverage is layered and heterogeneous. When the amount of Arabic gum added is large, the compound beverage will be more viscous. As shown in FIG. 8, Arabic gum with an addition of

0.4% is the optimum.

[73] 7. Results of orthogonal experiment on compounded Moringa seeds and

Ganoderma lucidum fermentative liquid

[74] A stability study on the compound beverage of Moringa seeds and a Ganoderma lucidum fermentative liquid is finally carried out by an orthogonal experiment with three factors: malic acid, xylitol and Arabic gum. According to results of the orthogonal experiment, malic acid has the greatest impact on the taste of the compound beverage, followed by Arabic gum, and finally xylitol. An optimum combination of these additives is 0.03% malic acid, 4.00% xylitol and 0.5% Arabic gum.

[75] 8. Sensory evaluation results

[76] The compound beverage of the Moringa seed liquid and the Ganoderma lucidum fermentative liquid is creamy white with a faint fragrance of Moringa seeds, the beverage is homogeneous and tastes sweet and sour.

Claims (6)

Conclusions 1. Ganoderma lucidum fermented liquid food, which includes a Ganoderma lucidum fermented liquid and a Moringa seminal fluid. The fermented liquid food of Ganoderma lucidum according to claim 1, wherein a ratio of the fermented liquid of Ganoderma lucidum to the Moringa seminal fluid is 1:1-5. The fermented liquid food of Ganoderma lucidum according to claim 1, further comprising an acidulant, a sweetener and a stabilizer. The fermented liquid food of Ganoderma lucidum according to claim 3, wherein the acidifying agent is malic acid. The fermented liquid food of Ganoderma lucidum according to claim 3, wherein the sweetener is xylitol. The fermented liquid food of Ganoderma lucidum according to claim 3, wherein the stabilizer is gum arabic.