팽나무버섯 액체 종균 배양시 이산화탄소 농도와 균사 생장량 변화

심규광,유영진,구창덕,김영석,김명곤 한국버섯학회 2012 한국버섯학회지 Vol.10 No.1

In this study, to produce Flammulina velutipes mushroom liquid spawn efficiently and effectively the effects of explosive aeration (supplying air with tiny bubbles) of the liquid culture medium on carbon dioxide concentration and residual sugar content in the medium on carbon dioxide concentration and residual sugar contentin the medium were measured. Carbon dioxide concentrations were measured at the outlet of the incubator. On the third day the explosive aeration greatly increased mycelial growth of the liquid spawn, and carbon dioxide concentration also greatly increased but decreased after 5 days. Free sugar contents in the liquid culture consistantly decreased up to 7 days and thereafter was not detected. The weight of the mycelia were maintained similar levels after 3 days. Total nitrogen content in the liquid medium constantly decreased during the 11days of explosive aeration. The content of free sugars in 7 days of culture was the lowest level, thus the inoculum incubated for 6~7 days was thought to be the most effective. Carbon dioxide concentration measurement at the outlet of the container during the liquid spawn incubation required low cost but was efficient to estimate the degree of mycelial growth to be used as a simple indicator.

팽나무버섯의 균사배양 중 배양기 내부 통기성 개선

심규광,유영진,구창덕,김영석,김명곤 한국버섯학회 2012 한국버섯학회지 Vol.10 No.1

Ventilation effects of bottles(1,100㎖) for culturing Flammulina velutipes on its mycelial growth and mushroom production were investigated. The degree of ventilation were controlled with hole positions, upper and under, and hole sizes in the bottle lids. The ventilation effects were measured with the contents of carbon dioxide, free sugars, chitin, moisture in the bottles and with the amount of produced mushrooms from the bottles. Carbon dioxide concentrations within the culturing bottles at exponential mycelial growth period vertex were relatively high in the bottles with lids without both a sponge and an aeration hole, and in those with a smaller hole. Free sugar contents in the mycelia were the highest in those with a 47mm hole on both sides, and in those with 26~33mm holes only underside. Chitin content was the highest in those with a 26mm hole only underside. On the other hand, the lids with 42mm~47mm holes on the both sides greatly lost water and decreased the mushroom production. In conclusion, the most efficient ventilation hole sizes on the lids for bottle(1,100㎖) cultivation of Flammulina velutipes using 1,100㎖ polypropylene bottle were 19mm on both sides of the lid and 26mm on only underside. They produced more mushrooms than the control by 6~9 %.

팽나무버섯 액체 종균의 접종 전 오염 검사

심규광,유영진,구창덕,김명곤 한국버섯학회 2012 한국버섯학회지 Vol.10 No.1

In this study, whether Giemsa staining solution can accurately determine bacterial contamination of liquid spawn for Flammulina velutipes in a short period of time was investigated. Giemsa solution staining cells of blood, bone marrow, lymph node, malaria parasites, rickettsia et al. was prepared by dissolving basic methylene azul and methylene blue, and acidic eosine in methyl alcohol-glycerine. Supernatant samples of Flammulina velutipes liquid spawn cultured under explosive aeration were placed on a slide, mixed with Gimesa solution and examined with optical microscope after staining. In 40 to 60 seconds bacterial cells were distinguishable from soybean meal residual and hyphal cell fragments. Thus we conclude that microscopy using Gimesa staining solution is a quick, simple and accurate method for the mushroom growers to effectively use to detect bacterial contamination of the liquid spawn.

표고 액체종균 배양시 배지와 균사체의 양분변화

심규광,유영진,구창덕,김명곤 한국균학회 2014 韓國菌學會誌 Vol.42 No.2

Lentinula edodes liquid spawn growth under explosive aeration (supplying air with tiny bubbles) and soybean mealaddition to liquid culture medium were investigated in terms of mycelial growth and residual free sugar content. The twotreatments were effective for homogeneous culturing of mycelial spawn and for separating colonies during multiplication after anexponential growth period without limiting sustaining nitrogen nutrients. The mycelial growth and carbon dioxide concentrationwere greatest on the 13th day since the inoculation. At 12th day, however, free sugars were almost depleted in the upper part ofthe liquid medium. Total nitrogen content within precipitated mycelia was the highest at the 13th day. Chitin and sucrose contentsin the mycelia were the highest at the 18th day, but ergosterol content became highest at 22 days. These results suggest thatLentinula edodes liquid spawn is ready in 18 days after inoculation. 본 연구에서는 효율적으로 표고 액체종균을 생산하기 위하여, 액체배지에 폭기방법으로 산소를 공급하고, 대두박을 첨가하여 표고균사체 생장량과 잔존 배양액의 유리당함량을 조사하였다. 그 결과 폭기방법은 균질화된 표고균사가 생장하면서 서로 뭉치지 않고 균질하게 증식하고, 대두박 첨가는 대수기 이후에도 지효성 질소영양원이 공급되도록 하는 효과가 있었다. 배양 중 침전 균사체의 중량은13일째에 가장 많았으며, 배양용기의 배출구에서 이산화탄소 농도는 13일째에 가장 높았다. 상층 수용액에서 환원당은 폭기 12일째에 거의 소비되었다. 그리고 배양균사체에서 총질소(T-N)량은 폭기 배양 13일째에 최고 수준이며, 키틴함량과 5종의 유리당 중에서 수크로즈함량은 폭기 배양18일째에 가장 높았으나, 에르고스테롤 함량은 폭기 배양22일째에 가장 높았다. 이와 같은 결과를 종합적으로 판단할 경우 표고액체 종균으로서 사용 적기는 균사활력이 가장 왕성하고 균체량이 많은 배양 18일째로 판단되었다.

천연발효종 유래 효모(Saccharomyces cerevisiae GDO Y45)의 분리 및 동정

심규광 ( Kyu-kwang Shim ),강동오 ( Dong-oh Kang ) 전북대학교 농업과학기술연구소 2023 농업생명과학연구 Vol.54 No.2

Strong fermentative yeasts were isolated during the preparation of natural sourdough, a continuous fermentation sourdough for Korean wheat powder. Colonies selected through the dilution plate process strongly formed black color in YA medium containing esculine, are yeasts that do not decompose protein components (casein), and new yeasts with strong fermentation power were selected through several repetitive processes. The selected strains have a large diameter of colonies formed due to their rapid growth rate, are yeasts that degrade cellulose, and are a new yeasts that do not degrade protein components. We finally selected GDO No. Y36, Y40, and Y45 among yeasts with repeatedly good swelling and strong alcohol odor during our wheat fermentation process and little odor of Korean wheat powder. As a result of analyzing the three contaminants that cause odor during the manufacture of natural sourdough, all had strong proteolytic power and fast growth rate, and all of them were identified as Bacillus spp. The results of examining the fermentation power of our wheat by preparing an aerobic liquid culture starter by selected yeast are as follows. In the process of fermenting the yeast culture solution and Korean wheat by mixing the same amount, fermentation proceeds actively after the third fermentation of Korean wheat (day 3). From then on, if the swelling of the fermentation dough is removed bubble space when the maximum swelling is formed in the fermentation of Korean wheat, the maximum swelling is repeated after an average of 3.4 hours. In the third sourdough of our Korean wheat using aerobic cultures of new yeast, it was found to be fermented 2.8-3.9 times the base volume. Without commercial yeast, new yeast alone has good swelling and can be baked, and all three strains were identified as Saccharomyces cerevisiae. S. cerevisiae GDO No. Y36, Y40, and Y45 isolated in this study are useful methods for the manufacture of natural sourdough of baking by manufacturing middle and large scale volume aerobic liquid culture starters. Among them, strains of S. cerevisiae GDO No. Y45 yeast have been deposited in KCTC (KCTC15481BP).

표고 액체종균 배양시 배지와 균사체의 양분변화

심규광 ( Kyu Kwang Shim ),유영진 ( Young Jin Yoo ),구창덕 ( Chang Duck Koo ),김명곤 ( Myung Kon Kim ) 한국균학회 2014 Mycobiology Vol.42 No.2

본 연구에서는 효율적으로 표고 액체종균을 생산하기 위하여, 액체배지에 폭기방법으로 산소를 공급하고, 대두박을 첨가하여 표고균사체 생장량과 잔존 배양액의 유리당함량을 조사하였다. 그 결과 폭기방법은 균질화된 표고균사가 생장하면서 서로 뭉치지 않고 균질하게 증식하고, 대두박 첨가는 대수기 이후에도 지효성 질소영양원이 공급 되도록 하는 효과가 있었다. 배양 중 침전 균사체의 중량은 13일째에 가장 많았으며, 배양용기의 배출구에서 이산화탄소 농도는 13일째에 가장 높았다. 상층 수용액에서 환원당은 폭기 12일째에 거의 소비되었다. 그리고 배양균사체에서 총질소(T-N)량은 폭기 배양 13일째에 최고 수준이며, 키틴함량과 5종의 유리당 중에서 수크로즈함량은 폭기 배양 18일째에 가장 높았으나, 에르고스테롤 함량은 폭기 배양 22일째에 가장 높았다. 이와 같은 결과를 종합적으로 판단 할 경우 표고액체 종균으로서 사용 적기는 균사활력이 가장 왕성하고 균체량이 많은 배양 18일째로 판단되었다. Lentinula edodes liquid spawn growth under explosive aeration (supplying air with tiny bubbles) and soybean meal addition to liquid culture medium were investigated in terms of mycelial growth and residual free sugar content. The two treatments were effective for homogeneous culturing of mycelial spawn and for separating colonies during multiplication after an exponential growth period without limiting sustaining nitrogen nutrients. The mycelial growth and carbon dioxide concentration were greatest on the 13th day since the inoculation. At 12th day, however, free sugars were almost depleted in the upper part of the liquid medium. Total nitrogen content within precipitated mycelia was the highest at the 13th day. Chitin and sucrose contents in the mycelia were the highest at the 18th day, but ergosterol content became highest at 22 days. These results suggest that Lentinula edodes liquid spawn is ready in 18 days after inoculation.

느타리의 균사 배양 중 배양기 내부 통기성 개선

유영진,심규광,구창덕,김명곤 한국버섯학회 2012 한국버섯학회지 Vol.10 No.2

The plastic culture bottle cap types and accumulated concentration of carbon dioxide, media humidity in the process of medium culture, chitin content and yield were observed in Pleurotus ostreatus 850ml bottle in Iksan, Jeollabuk-do, Korea, during 2011. The concentration of carbon dioxide in the process of medium culture was the highest after 8~9 days cultivation irrespective of cap sizes and types. The accumulated concentration of carbon dioxide in size cap of 29~41mm was 6.5~4.0% in the upper-under perforation hole of cap and 9.0~6.5% in the under perforation hole of cap. The upper-under 23~33mm perforation hole and under 29mm perforation hole of caps in the 850ml bottle were best condition for cultivation of mushroom and increased fruit body, 15.8~21.2% and 20%, respectively. However, the upper-under & under 41mm perforation hole of fruit body were decreased 60.7% and 23.6%, respectively. Also it was weak, lose vitality and the lower of biologically activity substance because the upper medium humidity was too dry.

느타리(1100㎖병)의 균사 배양 중 배양기 내부 통기성 개선

유영진,심규광,구창덕,김명곤 한국버섯학회 2012 한국버섯학회지 Vol.10 No.2

The plastic culture bottle cap types and accumulated concentration of carbon dioxide, media humidity in the process of medium culture and yield were observed in Pleurotus ostreatus 1,100ml bottle in Iksan, Jeollabuk-do, Korea, during 2011. The concentration of carbon dioxide in the process of medium culture was the highest after 6~11days cultivation irrespective of cap sizes and types. The upperunder 19~38mm perforation hole and under 26~47mm perforation hole of caps in the 1,100ml bottle were best condition for cultivation of mushroom and increased fruit body, 11.4~23.8% and 6.5~17.9%, respectively. However, the upper- under 33mm perforation hole of fruit body were decreased 23.8%. Also it was weak, lose vitality and the lower of biologically activity substance because the upper medium humidity was too dry.

Cyclodextrin 분해효소의 정제 및 그 특성

문영희,김용휘,심규광 한국농화학회 1990 Applied Biological Chemistry (Appl Biol Chem) Vol.33 No.1

Cyclodextrin hydrolase from Bacallus stearodhermophilus KFCC 21203 was purified and the properties of the purified enzyme were investigated. The enzyme was purified 15 folds with 77% recovery by ammonium sulfate fractionation, DEAE-cellulose chromatography, and Ultro AcA 34 gel filtration. The specific activity and the molecular weight of the enzyme were 1.30 units/㎎ protein and about 29,500, respectively. The maximum activity of the enzyme was shown at 55℃ and pH 5.5. However, stable temperature and pH were 40 ℃ and 5.0∼8.0, respectively. The Km value for γ-cyclodextrin was 3.78 × 10^(-3) M. The degradation activity of the enzyme was selectively high for γ-cyclodextrin, and very low for β-cyclodextrin, but not for α-cyclodextrin. The decomposed products of γ-cyclodextrin were mainly glucose and maltose, and a little mlatotriose. The activity of the enzyme was very high for amylose, potato starch, corn starch, amylopectin and maltooligomer, and relatively high for glycogen and dextrin. The decomposed products of them were mainly glucose and maltose.

아그배 Peroxidase의 정제 및 특성

양희천(Hee-Cheon Yang),손희숙(Hee-Suk Son),심규광(Kyu-Kwang Shim),오찬호(Chan-Ho Oh),최동성(Dong-Seong Choi) 한국식품영양과학회 1992 한국식품영양과학회지 Vol.21 No.1

아그배로 부터 아세톤 침전, DEAE-cellulose 칼럼 크로마토그라피, Ultro-AcA 54겔여과의 과정을 거쳐 peroxidase를 분리 정제하고, 그의 특성을 조사하였다. 아그배peroxidase의 반응 최적 pH는 4.5, 반응 최적 온도는 80℃이었고, 30℃ 이하의 온도와 pH 5.0에서 안정하였으며, 80℃에서 15분간 보존했을 때 거의 불활성화되었다. OPDA에 높은 활성을 나타내었으나, phenol류 기질에 대해서는 약간의 활성을 나타내었으며, OPDA와 H₂O₂에 대한 Km치는 각각 1. 65mM, 7.97mM이었다. 아그배 peroxidase에 대한 저해작용은 L-ascorbic acid와 sodium L-ascorbate가 가장 컸고, 금속이온 중 Mn^(2+)만이 5mM 농도에서 효소의 활성을 증가시켰다. Peroxidase in the fruit of Malus sieboldii (Regel) Rehder was partially purified by DEAE-cellulose column chromatography and Ultro-AcA 54 gel filtration. The optimum pH of peroxidase was 4.5 and optimum temperature was 80℃. The enzyme was stable at pH 5.0 and below 30℃, and inactivated by heat treatment at 80℃ for 15min. In the presence of 30mM H₂O₂ Km value on o-phenylenediamine as substrate was 1.65mM, and in the presence of 10mM o-phenylenediamine Km value on H₂O₂ was 7.97mM. L-Ascorbic acid and sodium L-ascorbate greatly inhibited the enzyme activity and among several metal ions Mn^(2+) only increased the activity at 5mM.