Rejuvenation method of special ganoderma lucidum strain for liquid fermentation

Abstract

The invention discloses a rejuvenation method of a special ganoderma lucidum strain for liquid fermentation. The method comprises the following steps: (1) shake-flask culture and screening: transferring activated parent ganoderma lucidum mycelia into a shake-flask for culture, and screening out shake-flask fermentation liquor of which the biological activity index is equivalent to that of the parent for later use; (2) plate culture and screening: re-inoculating the fermentation liquor which is obtained by shake flask screening and has the same biological activity index as the parent into a plate for culture, and selecting out the plate with high growth speed for later use; and (3) shake-flask culture verification: transferring the ganoderma lucidum mycelia cultured on the standby flat plate into a shake flask for subculture verification, wherein the flat plate strain used for shake-flask fermentation liquor with the biological activity index equivalent to that of a parent and stable subculture is the rejuvenated ganoderma lucidum strain. By applying the rejuvenation method disclosed by the invention, the problems of strain degradation, weak strain activity and poor strain quality caused in the passage process of liquid culture of the ganoderma lucidum strain can be effectively solved, and a strain guarantee is provided for liquid large-scale sustainable fermentation production of the ganoderma lucidum strain.

Description

technical field

[0001] The invention relates to the technical field of microbial strain breeding, in particular to a method for rejuvenating ganoderma strains specially used for liquid fermentation. Background technique

[0002] Ganoderma lucidum is a fungus of the genus Ganoderma in the family Polyporaceae of Basidiomycetes, and it is one of the rare medicinal fungi in my country. First recorded in the "Shen Nong's Materia Medica" in the Han Dynasty, it has the functions of strengthening the body, nourishing and strengthening, prolonging life, and maintaining the balance of the body. In recent years, Ganoderma lucidum has been deeply studied by many domestic and foreign scholars. At present, there are three ways to obtain the active ingredients of Ganoderma lucidum, one is wild fruiting bodies or artificially cultivated fruiting bodies, the other is Ganoderma lucidum spores, and the third is fermented mycelium and fermentation liquid. Compared with the artificial cultivat...

Examples

Embodiment 1

[0050] Parent Ganoderma lucidum strain: Ganoderma lucidum G0119 (provided and preserved by the Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, preservation number: HunongLingzhi No.1), which has been published in [Biotechnology and BioprocessEngineering 2014 Volume 19 Issue 4 727-732].

[0051] The Ganoderma lucidum strain after rejuvenation by this method, as shown in table 1 result, before rejuvenation, mycelia dry weight yield (15.12 ± 0.21) g / L, substrate utilization rate (64.62 ± 2.02)%, plate growth rate (6.39 ±0.30) mm / d, mycelium dry weight yield after rejuvenation (15.40±0.18) g / L, substrate utilization rate (66.11±1.60)%, plate growth rate (6.42±0.21) mm / d. The mycelium yield and substrate glucose utilization rate were stable before and after rejuvenation, and the fluctuation range of 12 consecutive passages was less than 8%.

Embodiment 2

[0053] Parent Ganoderma lucidum strain: G.lucidum G0017 (provided and preserved by the Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, preservation number: G.lucidum G0017), has been published in [International Journal of Medicinal Mushrooms 2021 Volume 23 Issue 3 Issue 43-53] public.

[0054] The Ganoderma lucidum strain rejuvenated by this method, as shown in table 1 result, mycelia dry weight yield (13.28 ± 0.26) g / L before rejuvenation, substrate utilization rate (73.33 ± 2.21) %, plate growth rate (5.89 ± 0.26) g / L 0.08) mm / d, mycelium dry weight yield after rejuvenation (14.02±0.24) g / L, substrate utilization rate (78.05±2.32)%, plate growth rate (5.99±0.06) mm / d. The mycelium yield and substrate glucose utilization rate were stable before and after rejuvenation, and the fluctuation range of 14 consecutive passages was less than 9%.

Embodiment 3

[0056]Parental Ganoderma lucidum strain: G.lucidum (provided and preserved by the Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, preservation number longzhi no.2), which has been published in [Journal of Edible Fungi, Vol. 27, No. 2, 2020, 84-91].

[0057] The Ganoderma lucidum strain rejuvenated by this method, as shown in table 1 result, mycelia dry weight yield (12.25 ± 0.12) g / L before rejuvenation, substrate utilization rate (73.33 ± 2.2) %, plate growth rate (5.53 ± 0.12) g / L 0.30) mm / d, mycelium dry weight yield after rejuvenation (12.98±0.24) g / L, substrate utilization rate (77.65±2.38)%, plate growth rate (5.73±0.18) mm / d. Before and after rejuvenation, mycelium yield and substrate glucose utilization were stable, and the fluctuation range of 11 consecutive passages was less than 7%.

[0058] In Examples 1-3, the comparison of the biological activity indexes before and after the rejuvenation of the ganoderma lucidum strain is shown in the follow...

technical field

[0001] The invention relates to the technical field of microbial strain breeding, in particular to a method for rejuvenating ganoderma strains specially used for liquid fermentation. Background technique

[0002] Ganoderma lucidum is a fungus of the genus Ganoderma in the family Polyporaceae of Basidiomycetes, and it is one of the rare medicinal fungi in my country. First recorded in the "Shen Nong's Materia Medica" in the Han Dynasty, it has the functions of strengthening the body, nourishing and strengthening, prolonging life, and maintaining the balance of the body. In recent years, Ganoderma lucidum has been deeply studied by many domestic and foreign scholars. At present, there are three ways to obtain the active ingredients of Ganoderma lucidum, one is wild fruiting bodies or artificially cultivated fruiting bodies, the other is Ganoderma lucidum spores, and the third is fermented mycelium and fermentation liquid. Compared with the artificial cultivat...

Examples

Embodiment 1

[0050] Parent Ganoderma lucidum strain: Ganoderma lucidum G0119 (provided and preserved by the Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, preservation number: HunongLingzhi No.1), which has been published in [Biotechnology and BioprocessEngineering 2014 Volume 19 Issue 4 727-732].

[0051] The Ganoderma lucidum strain after rejuvenation by this method, as shown in table 1 result, before rejuvenation, mycelia dry weight yield (15.12 ± 0.21) g / L, substrate utilization rate (64.62 ± 2.02)%, plate growth rate (6.39 ±0.30) mm / d, mycelium dry weight yield after rejuvenation (15.40±0.18) g / L, substrate utilization rate (66.11±1.60)%, plate growth rate (6.42±0.21) mm / d. The mycelium yield and substrate glucose utilization rate were stable before and after rejuvenation, and the fluctuation range of 12 consecutive passages was less than 8%.

Embodiment 2

[0053] Parent Ganoderma lucidum strain: G.lucidum G0017 (provided and preserved by the Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, preservation number: G.lucidum G0017), has been published in [International Journal of Medicinal Mushrooms 2021 Volume 23 Issue 3 Issue 43-53] public.

[0054] The Ganoderma lucidum strain rejuvenated by this method, as shown in table 1 result, mycelia dry weight yield (13.28 ± 0.26) g / L before rejuvenation, substrate utilization rate (73.33 ± 2.21) %, plate growth rate (5.89 ± 0.26) g / L 0.08) mm / d, mycelium dry weight yield after rejuvenation (14.02±0.24) g / L, substrate utilization rate (78.05±2.32)%, plate growth rate (5.99±0.06) mm / d. The mycelium yield and substrate glucose utilization rate were stable before and after rejuvenation, and the fluctuation range of 14 consecutive passages was less than 9%.

Embodiment 3

[0056]Parental Ganoderma lucidum strain: G.lucidum (provided and preserved by the Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, preservation number longzhi no.2), which has been published in [Journal of Edible Fungi, Vol. 27, No. 2, 2020, 84-91].

[0057] The Ganoderma lucidum strain rejuvenated by this method, as shown in table 1 result, mycelia dry weight yield (12.25 ± 0.12) g / L before rejuvenation, substrate utilization rate (73.33 ± 2.2) %, plate growth rate (5.53 ± 0.12) g / L 0.30) mm / d, mycelium dry weight yield after rejuvenation (12.98±0.24) g / L, substrate utilization rate (77.65±2.38)%, plate growth rate (5.73±0.18) mm / d. Before and after rejuvenation, mycelium yield and substrate glucose utilization were stable, and the fluctuation range of 11 consecutive passages was less than 7%.

[0058] In Examples 1-3, the comparison of the biological activity indexes before and after the rejuvenation of the ganoderma lucidum strain is shown in the follow...

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