• Title/Summary/Keyword: Strain analysis

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Purification and Characterization of a Lipolytic Enzyme Produced by Aeromonas hydrophila PL43 (Aeromonas hydrophila PL43이 생산하는 지질분해 효소의 정제 및 특성)

  • Kim, Yong-Woo;Hong, Sung Wook;Chung, Kun Sub
    • Microbiology and Biotechnology Letters
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    • v.44 no.2
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    • pp.130-139
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    • 2016
  • A bacterial strain, producing an excellent lipolytic enzyme, was isolated from the intestinal tracts of an earthworm (Eisenia fetida). The strain was identified as Aeromonas hydrophila by phenotypic, chemotaxonomic characteristics and 16S ribosomal DNA analysis, and was designated as Aeromona hydrophila PL43. The lipolytic enzyme from A. hydrophila PL43 was purified via 35−45% ammonium sulfate precipitation, DEAE-sepharose fast flow ion-exchange, and sephacryl S-300HR gel filtration chromatography. The yield of the purified enzyme was 3.7% and 2.5% of the total activity of crude extracts with p-nitrophenyl butyrate (pNPB) and p-nitrophenyl palmitate (pNPP) as substrates, respectively. The molecular weight of the purified enzyme was approximately 74 kDa using gel filtration, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and zymography. The optimal activity of purified enzyme was observed at 50℃ and pH 8.0 using pNPB, and 60℃ and pH 8.0 using pNPP. The purified enzyme was stable in the ranges 20− 60℃ and pH 7.0−10.0. The activity of purified enzyme was inhibited by PMSF, pepstatin A, Co2+, Cu2+, and Fe2+, but was recovered by metal chelating of EDTA. The Km and Vmax values of the purified enzyme were 1.07 mM and 7.27 mM/min using pNPB and 1.43 mM and 2.72 mM/min using pNPP, respectively.

The Optimal Production and Characteristics of an Alginate-degrading Enzyme from Vibrio sp. PKA 1003 (Vibrio sp. PKA 1003의 알긴산 분해 조효소 생산 최적 조건과 조효소의 특성)

  • Kim, Hyun-Jee;Kim, Koth-Bong-Woo-Ri;Kim, Dong-Hyun;SunWoo, Chan;Jung, Seul-A;Jeong, Da-Hyun;Jung, Hee-Ye;Lim, Sung-Mee;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.3
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    • pp.434-440
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    • 2013
  • This study was conducted to screen the characteristics and alginate-degrading activity of marine bacterium isolated from brown seaweed (Sargassum thunbergii). The results of 16S ribosomal RNA sequence analysis the strain the genus Vibrio and the strain was subsequently named Vibrio sp. PKA 1003. The optimum culture conditions for the growth of Vibrio sp. PKA 1003 were at pH 7, 3% NaCl, $25^{\circ}C$, and 6% alginic acid, with a 48-hour incubation time. A crude enzyme preparation from Vibrio sp. PKA 1003, showed its highest levels of alginate-degradation activity when cultured at pH 9, $30^{\circ}C$, and 6% alginic acid, with a 63-hour incubation time. Thin layer chromatography analyses confirmed that the crude enzyme released monomers or oligomers from sodium alginate, and results from trypsin treatment showed that the alginate degrading activity depends on this enzyme produced by Vibrio sp. PKA 1003. These results suggest that Vibrio sp. PKA 1003 and its alginate-degrading crude enzyme is useful for the production of alginate oligosaccharides.

Epidemiologic Investigation for the Etiology of an Epidemic Ocurred among Animals and Humans in an Isolated Island, Korea(I) (신안군(新安郡) 낙도(落島)에서 발생(發生)한 괴질(怪疾)의 원인(原因)에 관한 역학적(疫學的) 조사(調査)(I))

  • Kim, J.S.;Heo, Y.;Yoon, H.Y.;Lee, W.Y.
    • Journal of Preventive Medicine and Public Health
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    • v.22 no.2 s.26
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    • pp.290-301
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    • 1989
  • This is preliminary report on anthrax epidemic occurred in an island with about 100 residents. Since 1982 there had been sudden deaths among all kinds of domestic animals including cattle, dogs, ducks, chicken and goat but only a few among cats in an isolated island about three hours distance away by ferry boat from Mokpo city. From 1986 through 1988 nine human deaths and four patients occurred, which made the government intervene for investigation on June 25 1988. The epidemiological investigation consisted of interview survey and medical examination, medical record analysis, laboratory work to isolate the pathogens under the direction of hypothesis derived from the study and further confirmation of the pathogens by international institute. The summarized results are as followings: 1. According to the interview survey there were many deaths among domestic animals usually in cold and dry season such as January through March and September through November; 36 heads of cattle leaving one head, more than 40 hogs(all), hundreds of chicken leaving few alive, goats that had taken home from mountain and two or three cats out of around 40 had sudden deaths from 1982 till 1985, when the residents stopped to purchase and take them into the island anymore. Also there were eleven persons who had experienced the similar syndrome complex to those of admitted and expired patients and four of them revealed typical chest X-ray findings; from one of these four patients(Rho) B. anthracis is isolated. 2. Medical record on patients who had been admitted, showed common characteristics of the disease course. On admission they had either gastrointestinal or upper respiratory infection symptoms which invariably progressed to septicemic nature with pulmonary interstitial infiltration and mediastinal widening/bulging, and then to deadly acute respiratory distress syndrome. At the end stage chest X-ray revealed multiple bullous emphysema. One of another characteristics was oral ulceration with bleeding occurred in about 50% of the patients. Laboratory test results in common were leukocytosis with left shift and abnormal liver and kidney functions, particularly at the later stage of the illness. 3. Epidemiological characteristics was striking in that both mortality and incidence rates were high: the mortality rate was 8.7% average, male being three times higher than females but there was no distictive clustering by age group. The incidence rate for both sexes was 28.2% and there was no sex difference although a tendency of higher incidence among older ages was noticed. The highest mortality and incidence were observed in Won village where the first death of animal occurred and with the highest frequency among three villages of the island. 4. Among twelve bacilli species isolated from various specimens, two strains, one from patient and the other from soil where the recently died cow is hurried, were confirmed as B. anthracis by Pasteur Institute and CDC of USA(strain from soil). CDC reported that the strain did not produce capsule in bicarbonate media but reacted with the bacteriophage and one of five sera taken from the patients. Mode of transmission as well as incubation period of the agent has not been established yet, which needs further investigation in relation to the antigenic structure of the variant when it is confirmed.

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Characteristics and breeding of a new cultivar of Pleurotus ostreatus 'Soltari' (느타리 신품종 '솔타리'의 육성 및 자실체 특성)

  • Oh, Min-Ji;Shin, Pyung-Gyun;Oh, Youn-Lee;Jang, Kab-Yeul;Woo, Sung-I;Kong, Won-Sik
    • Journal of Mushroom
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    • v.14 no.4
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    • pp.202-206
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    • 2016
  • Oyster mushrooms are widely cultivated and consumed in Korea. P. ostreatus 'Suhan(ASI 2504)' is an ideal cultivar for mushroom farmers due to its dark pileus and thick stipe; however, as it is very sensitive to environmental conditions, an alternative cultivar is required. To develop a new cultivar, parental strains 'Suhan(ASI 2504)' and 'ASI 0665 (Heuktari)' were selected from various collected strains according to morphological characteristics. P. ostreatus 'Soltari' was developed by DiMon crossing between the dikaryotic strain 'Suhan' and the monokaryotic strain derived from 'Heuktari'. Thirty-eight of the 100 crossed strains were selected following analysis of mitochondrial genetic characteristics, and 'Soltari' was ultimately selected by continuous cultivation tests. The mitochondrial DNA profile of 'Soltari' was found to be the same as that of 'Heuktari, and a nuclear DNA profile of 'Soltari' was similar as those of the parental strains, 'Suhan' and 'Heuktari.' 'Soltari' mycelium grows adequately in moderate to high temperatures of $12-20^{\circ}C$, although its optimum temperature was found to be $30^{\circ}C$. Fruiting body production per 1.1-L cultivation bottle was approximately 158.8 g. Its stipe length and thickness were comparable to those of diameter and thickness were somewhat lower (42.72 vs. 51.33 mm and 18.18 vs. 22.46 mm, respectively). 'Soltari' was found to be more resistant to high $CO_2$ atmosphere than 'Suhan', and the color of the pileus of 'Soltari' was dark gray at high temperature. Therefore, it is suggested that this new cultivar 'Soltari' is a good alternative cultivar and will contribute to energy saving in oyster mushroom farms.

Characterization of ${\alpha}$-Galactosidase and ${\beta}$-Glucosidase by Weissella cibaria (Weissella cibaria가 생산하는${\alpha}$-Galactosidase 및 ${\beta}$-Glucosidase의 특성)

  • Hong, Sung-Wook;You, Lae-Kyun;Jung, Byung-Moon;Kim, Wan-Sik;Chung, Kun-Sub
    • Microbiology and Biotechnology Letters
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    • v.37 no.3
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    • pp.204-212
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    • 2009
  • A strain producing ${\alpha}$-galactosidase and ${\beta}$-glucosidase was isolated from Kimchi. The isolated strain was identified as Weissella cibaria by 16S rDNA analysis and designated as Weissella cibaria K-M1-4. The enzyme activity of ${\alpha}$-galactosidase and ${\beta}$-glucosidase reached the maximum in the soy medium at $37^{\circ}C$ for 24 hr. The enzymes were purified by ethanol fractionation, DEAE sepharose fast flow, and sephacryl S-100HR column chromatography. ${\alpha}$-Galactosidase specific activity was shown by 576 Units/mg protein and the yield was 3.5% of the total activity of crude extracts. ${\beta}$-glucosidase specific activity was shown by 480 Units/mg protein and the yield was 2.9% of the total activity of crude extracts. The optimum temperature for ${\alpha}$-galactosidase was $60^{\circ}C$ and 43% of its original activity remained when it was treated at $80^{\circ}C$ for 30 min. For ${\alpha}$-galactosidase shows the optimum pH of 8.0 and is fairly stable between pH5.0 and pH9.0. The enzyme activity was increased in the presence of $Fe^{2+}$ and $Cu^{2+}$. The value of Km and Vmax for the enzyme were 0.98 mM and $1.81{\mu}$mole/min, respectively. The ${\beta}$-glucosidase has the optimum temperature of $50^{\circ}C$ and 46% of its original activity remained when it was treated at $80^{\circ}C$ for 30min. Its optimum pH of 7.0 and is fairly stable between pH5.0 and pH9.0. The enzyme activity was increased in the presence of $Fe^{2+},\;Co^{2+}$ and $Cu^{2+}$. The value of Km and Vmax for the enzyme were 1.24 mM and $6.81{\mu}$mole/min, respectively.

Isolation and Mycelial Cultivation Submerged of Phellinus sp. (Phellinus sp.의 분리 및 균사체의 액체배양)

  • Kang, Tae-Su;Lee, Dong-Gi;Lee, Shin-Young
    • The Korean Journal of Mycology
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    • v.25 no.4 s.83
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    • pp.257-267
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    • 1997
  • Fruit bodies similar to the Phellinus sp. residing on the mulberry were collected at Yang-yang in Kang-won-do province and one strain of Phellinus sp. was isolated from the fruit bodies. For mass production of the isolated mycelia in a submerged culture, the culture conditions, medium composition, and the effect of various culture systems on the mycelial growth, were investigated. The morphological characteristics of the fruit body were as follows: covered with blackish to black and rough, lower surface with yellowish-brown to dull-brown and smooth, 5-7 cm thick and hard woody. Also, the pure cultured mycelia showed yellowish-brown color, capability of purplish-brown pigment production on the PDA plate media, no-formation of clamp-connection, much binding branch, and enzyme activities such as laccase, tyrosinase and peroxidase. Therefore, pure cultured strain was identified to be Phellinus sp. In the flask culture, the optimum culture conditions for the mycelial production were obtained after cultivation of 8 days at inoculum level of 5%(v/v), media volume of 70 mL, 150 rpm, initial pH 6, and temperature of $30^{\circ}C$. Optimum medium composition from the response surface analysis were determined to be glucose 12.12 g/L, sucrose 12.12 g/L, yeast extract 11.15 g/L, malt extract 11.15 g/L, $KH_2PO_4$ 0.855 g/L and $CaCl_2$ 0.855 g/L. The production of the mycelia after 4 and 8 days of cultivation was 1.95 and 9.89 g/L, respectively. The maximum specific growth rate and productivity were $0.020\;hr^{-1}$ and 1.25 g/L/day, respectively. Among the three different culture systems for the growth of mycelia, the maximum mycelial dry weight of 7.5 g/L was obtained after cultivation of 4 days in the air-lift fermentor under aeration rate of 2.5 vvm. The maximum specific growth rate and productivity were $0.033\;hr^{-1}$ and 1.9 g/L/day, respectively, which were about 1.7 and 4.2 times higher than those of flask culture.

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The effect of the revolution and forwarding speed of the rotary blade on the tilling power requirement (로우터리 경운(耕耘)날의 회전속도(回轉速度) 및 작업속도(作業速度)가 경운소요동력(耕耘所要動力)에 미치는 영향(影響))

  • Kwon, Soon Goo;Kim, Soung Rai
    • Korean Journal of Agricultural Science
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    • v.11 no.1
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    • pp.160-175
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    • 1984
  • This study was carried out to analyze the effects of the revolution and forwarding speed of the rotary blade and the edge curves which were $30^{\circ}$ and $40^{\circ}$, on the power requirement of rotary tillage. In this study, the revolutions of the rotary blade considered were 204, 243, 285, 360 rpm, and the forwarding speeds of the rotary system considered were 29.40cm/sec, 46.93em/sec. The power requirements of rotary blade were measured by a dynamic strain gage systems at the soil bin which was filled with artificial soil. The results of the study were summarized as follows: 1. The response surface analysis showed that the revolution and forwarding speed of the rotary shaft had an interacting influence on the torque requirement of the rotary blade. The mathematical model developed by the above was repersented as follow. $$T=a_0+a_1V+a_2R +a_3VR+a_4VR^2$$ where, $a_0=constant$ $a_1,\;a_2,\;a_3,\;a_4=coefficients$ V=forwarding speed of the rotary system. (em/sec) R=revolution of the rotary shaft. (rpm) T=tilling torque requirement. (kg-m) 2. When the maximum tilling torque requirement was analyzed, ${\partial}T/{\partial}R$ was decreased with the increasing revolution of rotary shaft, while ${\partial}T/{\partial}V$ was increased, which was minimum at 200~220 rpm. When the forwarding speeds were increased, ${\partial}T/{\partial}R$ was decreased with increasing rate. 3. When the mean tilling torque requirement was analyzed, ${\partial}T/{\partial}V$ was constant at 320~360 rpm and ${\partial}T/{\partial}R$ was decreased with increasing rate along with the increasing revolution of rotary shaft. 4. When the mean tilling torgue requirement per unit volume of soil was analyzed, ${\partial}T/{\partial}V$ was minimum at 270~300 rpm. ${\partial}T/{\partial}R$ for the forwarding speeds of 29.40cm/sec and 46.93cm/sec was same as that for 280~290 rpm. 5. Increasing the edge curves of the rotary blades, the tilling torque requirement was increased. But other studies showed that the smaller the edge curve, the more straw could be wrapped on blades which resulted in increasing torque requirements. Therefore, the edge curve of rotary blade should be considered for the future study.

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Effect of Trehalose on the Viability of Fluorescent Pseudomonas, Strain SSL3 (형광성 Pseudomonas, SSL3 균주의 생존율에 미치는 Trehalose의 효과)

  • Seong, Ki-Young;Ryu, Ok-Ran;Choi, Won-Yeol
    • Korean Journal of Soil Science and Fertilizer
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    • v.33 no.4
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    • pp.292-301
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    • 2000
  • To make a better use of the beneficial bacterial inoculants in the agricultural practice, dry forms of bacterial fertilizer or pesticides are prepared with carrier materials. During the drying process of bacterial inoculant, most of the cells face a severe osmotic pressure and dehydration, and die off. Our study describes the effect of osmoprotectants such as trigonelline and trehalose on the survival of bacterial cells in high salt concentration and drying conditions. A fluorescent Pseudomonas, strain SSL3, used in this study, could grow in high salt concentration of upto 5% but the cells could not overcome the growth retardation at over 7% of salt concentration. The addition of trigonelline, even on small amount, in liquid medium containing 4% NaCl was detrimental to the cell. However, the addition of trehalose of upto 10 mM to the liquid medium containing 4% NaCl, enhanced cell growth. The cell growth was retarded when 150mM trehalose was added to the medium. Upon dry formulation of cells, trehalose was added. And the dry cells were inoculated into the soil to determine the effect of osmoprotectants on the survival of the cells. The survival of the cells, both in wet or dry soil, was improved by the addition of trehalose during the dry cell formulation. The positive effect of trehalose on the cell survival at $-20^{\circ}C$ and $-70^{\circ}C$ was oven more pronounced. The FTIR (Fourier transformation infra-red) spectroscopic analysis showed that the change of the 2nd amide group was reduced by adding trehalose to the medium containing 4% NaCl. These results suggest that trehalose can protect the cell membrane from dryness or high concentration of salt, thereby diminishing the sudden change of the protein structure of the cell membrane and, as a consequence, improving the cell survival.

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Biaxial Strain Analysis of Various Fixation Models in Porcine Aortic and Pulmonary Valves (돼지 대동맥 판막과 폐동맥 판막의 고정 방법에 따른 양방향 압력-신장도의 비교분석)

  • Cho, Sung-Kyu;Kim, Yong-Jin;Kim, Soo-Hwan;Choi, Seung-Hwa
    • Journal of Chest Surgery
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    • v.42 no.5
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    • pp.566-575
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    • 2009
  • Background: The function of a bioprosthetic heart valve is determined largely by the material properties of the valve cusps. The uniaxial tensile test has been studied extensively. This type of testing, however, does not replicate the natural biaxial loading condition. The objective of the present study was to investigate the regional variability of the biaxial strain versus pressure relationship based on the types of fixation liquid models. Material and Method: Porcine aortic valves and pulmonary valves were assigned to three groups: the untreated fresh group, the fixed with glutaraldehyde (GA) group, and the glutaraldehyde with solvent (e.g., ethanol) group. For each group we measured the radial and circumferential stretch characteristics of the valve as a function of pressure change. Result: Radial direction elasticity of porcine aortic and pulmonary valves were better than circumferential direction elasticity in fresh, GA fixed and GA+solvent fixed groups (p=0.00). Radial and circumferential direction elasticity of pulmonary valves were better than aortic valves in GA fixed, and GA+solvent fixed groups (p=0.00). Radial and circumferential direction elasticity of aortic valves were decreased after GA and GA+solvent fixation(p=0.00), except for circumferential elasticity of GA+solvent fixed valves (p=0.785). The radial (p=0.137) and circumferential (p=0.785) direction of elasticity of aortic valves were not significantly different between GA fixed. and GA+solvent fixed groups. Radial (p=0.910) and circumferential (p=0.718) direction of elasticity of pulmonary valve also showed no significant difference between GA fixed and GA+solvent fixed groups. Conclusion: When fixing porcine valves with GA, adding a solvent does not cause a loss of mechanical properties, but, does not improve elasticity either. Radial direction elasticity of porcine aortic and pulmonary valves was better than circumferential direction elasticity.

Breeding and characterization of 'Creamy', a new interspecific hybrid between Pleurotus ferulae and P. tuoliensis (아위느타리와 백령느타리의 종간교잡 품종 '크리미'의 육성 및 특성)

  • Oh, Min-Ji;Shin, Pyung-Gyun;Lim, Ji-Hoon;Oh, Youn-Lee;Jang, Kab-Yeul;Kong, Won-Sik
    • Journal of Mushroom
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    • v.17 no.4
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    • pp.224-229
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    • 2019
  • The two most common mushroom species grown in Korea are pearl oyster mushroom (Pleurotus ostreatus) and king oyster mushroom (P. eryngii). In recent years, the production of king oyster mushroom greatly increased due to the automation of the cultivation facilities, and it became a major export mushroom owing to its excellent shelf life. However, the increase in the production of king oyster mushroom led to a decline in its market price; thus, necessitating the development of new mushroom species that could replace king oyster mushroom, to diversify the mushroom market for the benefit of both, the producers and the consumers. The Mushroom division at the National Institute of Horticultural & Herbal Science (NIHHS) reported the development of a new interspecific hybrid between P. ferulae and P. tuoliensis, referred to as 'Creamy.' Two parental strains KMCC00430 (Bisan2ho, P. ferulae) and KMCC00461 (P. tuoliensis) were selected based on the results of genetic resource analysis, and their monokaryons were collected. About 1,000 Mon-Mon crosses were performed and 73 of them were selected. Following repeated cultivation tests and strain analyses, we selected strain 7773, which had a bright creamy pileus and a thick straight stipe, and named it 'Creamy.' Optimum temperature for mycelial growth of Creamy was 25-30℃, and that for fruiting body growth was 16℃. The pileus, which had a brighter creamy color, was small in size with a diameter of 61.2 mm. Although it was cultivated in suboptimal conditions, such as low temperature and high CO2 concentration, Creamy was characterized by its straight and smooth stipe. Field production tests and further analyses indicated that the yield of Creamy was 5% higher than that of Baekhwang. It is expected that Creamy, the new interspecific hybrid with a bright creamy pileus and a pleasant flavor, will help create new opportunities for mushroom farmers and diversify the mushroom market.