Improved cordycepin production in a liquid surface culture of Cordyceps militaris isolated from wild strain

Abstract

A Cordyceps militaris NBRC 10352-3 strain that was isolated from C. militaris NBRC10352 produced 68 mg of cordycepin from 100 ml of medium, which was the highest level of cordycepin among 60 isolates from three C. militaris (NBRC 9787, 100741 and 103752) strains. Interestingly, a liquid surface culture of C. militaris NBRC 103752-3 produced 2-fold cordycepin to that in a submerged culture. Cordycepin production was significantly affected by specific surface area (SSA) in the liquid surface culture, and 120.9 mg of cordycepin was produced on SSA of 1.57 cm-1 (from 50 ml). The addition of glycine and adenine as an additive to its culture medium was optimized by an experimental design. When 6.75 g/l of adenine was added to the culture, 315.7 mg of cordycepin was produced from 50 ml medium, improving the cordycepin production by 4.7-fold. In this study, the production and productivity of cordycepin were significantly improved in C. militaris wild type by a single cell colony isolation and additives without adopting any mutational technologies. This C. militaris NBRC 10352-3 strain can be used as a new cordycepin-hyperproducing one, instead of a cordycepinhyperproducing mutant.

Metabolic comparison of aerial and submerged mycelia formed in the liquid surface culture of Cordyceps militaris

“…The consumption of phenylalanine suggested that it could be a candidate of additives to enhance the production of cordycepin. However, in our previous study, the addition of phenylalanine did not have any effect on cordycepin production in C. militaris (Sari, Suparmin, Kato, & Park, ).…”

“…This fungus was inoculated into an optimized culture medium (Sari et al, ) composed of 72.5 g/L yeast extract, 62.6 g/L glucose (pH 5.6) and Vogel's medium with 1/10 concentration containing 0.28 g/L sodium citrate dihydrate, 0.50 g/L KH 2 PO 4 , 0.20 g/L NH 4 NO 3 , 0.02 g/L MgSO 4 ·H 2 O, 0.01 g/L CaCl 2 2H 2 O, 0.46 × 10 −3 g/L citric acid, 0.50 × 10 −3 g/L ZnSO 4 , 0.1 × 10 −3 g/L Fe(NH) 4 (SO 4 ) 2 .6H 2 O, and 0.025 × 10 −3 g/L CuSO 4 5H 2 O and incubated at 25°C for 19 days in liquid surface culture.…”

Insight into cordycepin biosynthesis of Cordyceps militaris: Comparison between a liquid surface culture and a submerged culture through transcriptomic analysis

“…C . militaris was cultivated in an optimized production medium [ 15 ] composed of 72.5 g/L yeast extract, 62.6 g/L glucose (pH 5.6) and Vogel’s medium with 1/10 concentration containing 0.28 g/L sodium citrate dihydrate, 0.50 g/L KH 2 PO 4 , 0.20 g/L NH 4 NO 3 , 0.02 g/L MgSO 4 ·H 2 O, 0.01 g/L CaCl 2 ·2H 2 O, 0.46 × 10 −3 g/L citric acid, 0.50 × 10 −3 g/L ZnSO 4 , 0.1 × 10 −3 g/L Fe(NH) 4 (SO 4 ) 2 ·6H 2 O, and 0.025 × 10 −3 g/L CuSO 4 ·5H 2 O. One disk (1 cm diameter) of mycelia in PDA plates that had been cultured for 7 d was inoculated into 100 ml of the optimized production medium in a 500 ml Erlenmeyer flask and incubated at 25°C for 19 d in the LSC.…”

Effects of Cordycepin in Cordyceps militaris during Its Infection to Silkworm Larvae

“…Additionally, cordycepin shows insecticidal activity to Plutella xylostella (Lepidoptera: Plutellidae) and Trypanosoma evansi [ 9 , 10 ]. Large amounts of cordycepin are produced in medium from surface cultures of mycelia [ 11 , 12 ]. This culture method supports large-scale production using mycelial cultivation instead of the preparation of fruiting bodies.…”