Optimization of Oudemansiella raphanipes Liquid Fermentation Medium

doi:10.15933/j.cnki.1004-3268.2024.05.014

Abstract

Abstract: Oudemansiella raphanipes strain H1 was used as experimental material and single factor test，Plackett‑Burman design，maximum climb test and Box‑Benhnken design response surface method were used to optimize the liquid culture medium of strain H1 with mycelial dry weight as the main evaluation index,in order to improve the mycelium biomass of O.raphanipes liquid seeds，and provide the guarantee for the industrial large‑scale production of O.raphanipes.The results showed that the most suitable carbon source for the growth of O.raphanipes was corn meal and the most suitable nitrogen source was
soybean meal.When the rotating speed of shaker was 150 r/min and the culture temperature was 25℃，the formula of optimized liquid fermentation medium was 25.8 g/L glucose，26.0 g/L corn flour,1.0 g/L KH₂PO_4，1.5 g/L soybean meal，1.0 g/L yeast extract powder，0.4 g/L MgSO_4，0.01 mg/L VB_1，and under this culture condition，the measured value of mycelium dry weight was 23.2 g/L.The culture cycle was shortened by 3 d，and the mycelial activity was higher compared with conventional medium.

Key words: Oudemansiella raphanipes, Plackett?Burman experiment, Response surface method, Liquid spawn, Fermentation medium

摘要： 以卵孢长根菇菌株H1为试材，以菌丝生物量为主要评价指标，在碳氮源单因素试验的基础上，采用Plackett-Burman设计对影响卵孢长根菇液体发酵培养生物量的主效因素进行筛选，通过最陡爬坡试验及Box-Benhnken响应面法对卵孢长根菇液体发酵培养基成分进行优化，以期获取优良液体菌种，为工厂化大规模生产卵孢长根菇提供保障。结果表明，适合卵孢长根菇的最适碳源为玉米粉，最适氮源为豆粕粉。当摇床转速150 r/min、培养温度为25 ℃时，优化后卵孢长根菇液体发酵培养基成分为葡萄糖25.8 g/L、玉米粉26.0 g/L、KH₂PO₄ 1.0 g/L、豆粕粉1.5 g/L、酵母粉1.0 g/L、MgSO₄ 0.4 g/L、维生素B₁（VB₁）0.01 mg/L，其卵孢长根菇菌丝生物量可达23.2 g/L。与常规培养基对比，培养周期缩短3 d，菌丝活力更高。

关键词: 卵孢长根菇, Plackett-Burman试验, 响应面, 液体菌种, 发酵培养基

CLC Number:

S646

LIU Guoli, CHEN Xun, XIAO Jun, MA Xiaoying, ZHANG Min, ZHANG Peng, ZHANG Man, GONG Na. Optimization of Oudemansiella raphanipes Liquid Fermentation Medium[J]. Journal of Henan Agricultural Sciences, 2024, 53(5): 133-140.

刘国丽, 陈珣, 肖军, 马晓颖, 张敏, 张鹏, 张曼, 龚娜. 卵孢长根菇液体发酵培养基配方优化[J]. 河南农业科学, 2024, 53(5): 133-140.

References

［1］黄书文，刘叶高，张文华，等.长根菇单孢杂交育种试验初报［J］.食药用菌，2021，29（6）：523-526.

HUANG S W，LIU Y G，ZHANG W H，et al. Preliminary report on single spore hybridization breeding of Oudemansiella radicata ［J］. Edible and Medicinal Mushrooms，2021，29（6）：523-526.

［2］苗晶囡，董嘉木，刘树民，等.黑皮鸡枞菌多糖超声辅助提取条件优化及抗氧化活性研究［J］.食品安全质量检测学报，2023，14（12）：109-116.

MIAO J N，DONG J M，LIU S M，et al. Ultrasonic-assisted extraction condition optimization and antioxidant activity of polysaccharides from Oudemansiella raphanipes［J］.Journal of Food Safety & Quality，2023，14（12）：109-116.

［3］WANG Y F，JIA J X，REN X J，et al. Extraction，preliminary characterization and in vitro antioxidant activity of polysaccharides from Oudemansiella radicata mushroom［J］. International Journal of Biological Macromolecules：Structure，Function and Interactions， 2018，120（Pta2）：1760-1769.

［4］安晓雯，王彦立，杨子怡，等.黑皮鸡枞菌营养与质构特性分析及其抗氧化活性评价［J］.食品工业科技，2021， 42（5）：236-242，249.

AN X W，WANG Y L，YANG Z Y，et al. Analysis of nutritional and textural properties and antioxidant activity evaluation of Termitornyces albuminosus［J］.Science and Technology of Food Industry，2021，42（5）：236-242，249.

［5］TSANTRIZOS Y S，YANG X S，MCCLORY A. Studies on the biosynthesis of the fungal metabolite oudenone.2.Synthesis and enzymatic cyclization of an alpha-diketone，open-chain precursor into oudenone in cultures of Oudemansiella radicata［J］.Journal of Organic Chemistry，1999，64（18）：6609-6614.

［6］万鲁长，李晓博，赵敬聪，等.北方地区长根菇大棚地栽周年生产标准化技术［J］.食药用菌，2019，27（2）： 135-138.

WAN L Z，LI X B，ZHAO J C，et al.Annual production technique of Oudemansiella radicata in greenhouses in northern areas of China［J］. Edible and Medicinal Mushrooms，2019，27（2）：135-138.

［7］肖自添，凌宏通，曾令贤，等.卵孢长根菇（黑皮鸡枞菌）工厂化免覆土周年高效栽培技术［J］.中国蔬菜，2021 （6）：112-114.

XIAO Z T，LING H T，ZENG L X，et al.High-efficiency cultivation techniques of Oudemansiella raphanipes without covering soil in factory for the whole year［J］.China Vegetables，2021（6）：112-114.

［8］王涛，谭琦，李玉，等.杏鲍菇液体菌种应用工艺参数的优化［J］.上海农业学报，2019，35（1）：33-37.

WANG T，TAN Q，LI Y，et al. Optimization of process parameters for application of liquid spawn of Pleurotus eryngii［J］.Acta Agriculturae Shanghai，2019，35（1）： 33-37.

［9］姚强，宫志远，司洪宇，等.工厂化生产金针菇液体菌种配方开发及应用［J］.山东农业科学，2020，52（11）： 134-137.

YAO Q，GONG Z Y，SI H Y，et al. Formula development and application of liquid strain of Flammulina velutipes in factory production［J］.Shandong Agricultural Sciences， 2020，52（11）：134-137.

［10］周超，马银鹏，包旭翔，等.响应面法优化黑木耳液体菌种发酵配方及条件［J］.山西农业科学，2023，51 （6）：653-662.

ZHOU C，MA Y P，BAO X X，et al.Optimization of fermentation formula and conditions of Auricularia heimuer liquid strain by response surface methodology［J］.Journal of Shanxi Agricultural Sciences，2023，51 （6）：653-662.

［11］冯伟林，金群力，叶晓菊，等.黑木耳液体菌种的制备工艺及质量标准［J］.食药用菌，2022，30（1）：78-80.

FENG W L，JIN Q L，YE X J，et al. Preparation technology and quality standard of liquid spawn of Auricularia auriculata［J］.Edible and Medicinal Mushrooms，2022，30（1）：78-80.

［12］周婷婷，周荣翔，张瑶，等.杏鲍菇液体菌种的细胞固定化技术及其保藏条件研究［J］.河南农业科学， 2022，51（3）：118-126.

ZHOU T T，ZHOU R X，ZHANG Y，et al.Study on cell immobilization technology and storage conditions of liquid strains of Pleurotus eryngii［J］. Journal of Henan Agricultural Sciences，2022，51（3）：118-126.

［13］王宇星.香菇液体菌种高效生产及应用栽培研究［D］.上海：上海海洋大学，2022.

WANG Y X. Study on efficient production and applied to cultivation of Lentinula edodes liquid spawn［D］.Shanghai：Shanghai Ocean University，2022.

［14］刘运伟，郭兴.黑皮鸡枞菌液体培养基配方筛选［J］.中国林副特产，2020（4）：23-25.

LIU Y W，GUO X. Formula screening of liquid culture medium for Oudemansiella radicata［J］.Forest by Product and Speciality in China，2020（4）：23-25.

［15］鲁铁，陈怀中，李一，等.响应面优化卵孢小奥德蘑的液体菌种发酵工艺［J］.北方园艺，2020（22）： 116-121.

LU T，CHEN H Z，LI Y，et al. Optimization of liquid spawn fermentation technology of Hymenopellis raphanipes by response surface methodology［J］.Northern Horticulture，2020（22）：116-121.

［16］徐宁，冯立国，王春晖，等.黑皮鸡枞液体菌种发酵罐培养条件的优化［J］.南方农业学报，2019，50（2）： 344-349.

XU N，FENG L G，WANG C H，et al.Optimization of culture conditions of fermentation technology for Oudemansiella raphanipes liquid strain fermenter［J］. Journal of Southern Agriculture，2019，50（2）：344-349.

［17］张昊月，郭正彦，吕志堂，等.应用响应面法优化发酵培养基提高达托霉素产量［J］.微生物学通报，2021， 48（1）：113-122.

ZHANG H Y，GUO Z Y，LYU Z T，et al.Medium optimization with the response surface methodology to improve daptomycin production ［J］. Microbiology China，2021，48（1）：113-122.

［18］冀宏，梁永军，柳宁，等.杏鲍菇液体菌种质量评价及标准研究初探［J］.北方园艺，2016（6）：128-132. JI H， LIANG Y J，LIU N，et al. Research on quality evaluation and standard system of Pleurotus eryngii liquid spawn［J］.Northern Horticulture，2016（6）： 128-132.

［19］赵刚，顾佳丽，励建荣.Plackett-Burman设计及响应面法优化芹菜中总黄酮的超声提取工艺［J］.中国食品学报，2017，17（8）：147-155.

ZHAO G，GU J L，LI J R. Optimization of ultrasonic extraction conditions of total flavonoids from Apium graveolens by Plackett-Burman design and response surface analysis［J］.Journal of Chinese Institute of Food Science and Technology，2017，17（8）：147-155.

［20］谭强飞，王文沛，程志虹，等.草菇液体菌种培养基优化及与固体菌种栽培对比［J］.食品与发酵工业， 2023，49（12）：85-91.

TAN Q F，WANG W P，CHENG Z H，et al.Optimization of Volvariella volvacea liquid culture medium and its cultivation comparison with solid starter［J］.Food and Fermentation Industries，2023，49 （12）：85-91.

［21］代俊杰，黎智文，李晓.响应面法优化羊肚菌栽培种培养基［J］.分子植物育种，2018，16（24）：8162-8167.

DAI J J，LI Z W，LI X. Optimization of Morchella cultivars medium by response surface methodology［J］.Molecular Plant Breeding，2018，16（24）：8162-8167.

［22］魏姣.黑皮鸡枞菌种培养基优化及新型栽培基质研究［D］.长沙：湖南农业大学，2020.

WEI J. Study on the optimization of fir culture medium and the new cultivation matrix of Oudemansiella raphanipes［D］.Changsha： Hunan Agricultural University，2020.

［23］黄静，李鹏，和耀威，等.响应曲面法优化黑皮鸡枞液体配方研究［J］.安徽农业科学，2023，51（13）：49-52. HUANG J，LI P，HE Y W，et al. Study on optimization of liquid formula of Oudemansiella radicata by response surface method［J］.Journal of Anhui Agricultural Sciences，2023，51（13）：49-52.

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