• Title/Summary/Keyword: bacterial production

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Studies on Growth Stimulation of Lactic Starter (유산균(乳酸菌) Starter의 생육촉진(生育促進)에 관(關)한 연구(硏究))

  • Kim, Jong Woo;Lee, Young Soo
    • Korean Journal of Agricultural Science
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    • v.19 no.1
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    • pp.40-50
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    • 1992
  • This experiment was carried out to stimulate lactic starter culture for yoghurt manufacturing. A each of 1.5% of Bios 2000, CR starter medium, and Yeast extract were added to bulk medium, acidity, pH and changes in the number of lactic acid bacteria were investigated at, intervals of two hours for Lactobacillus bulgaricus and four hours for Streptococus thermophilus and Lactobacillus casei. The results obtained were summarized as follows. 1. The acidity of control arrived at 0.99% after 16 hours of incubation during the incubation of Lactobacillus bulgaricus. Whereas that of CR starter medium reached 1.00% at 12 hours of incubation. Yeast extract, 1.12% at 12 hours, and Bios 2000 reached 0.97% at 10 hours respectively, Thus, Bios 2000 showed the fastest rate of acid production. 2. When the acidity of experiment medium peaked on optimum levels. pH of control was 4.03 in 16 hours of incubation during the incubation of Lactobacillus bulgaricus. Whereas that of Bios 2000 reached 4.10 of Yeast extract reached 3.97 at 12 hours, and of CR starter medium reached 4.05 at 12 hours. 3. Lactic acid bacterial counts were $3.1{\times}10^{10}/ml$ after 16 hours of incubation during the incubation of lactobacillus bulgaricus, Whereas those of Bios 2000 reached $2.1{\times}10^{10}/ml$ at 10 hours, with the fastest stimulation of growth, The counts in CR starter medium were at $2.9{\times}10^{10}/ml$ at 12 hours, and Yeast extract were $3.8{\times}10^{10}/ml$ at 12 hours. 4. The acidity of control, CR starter medium, and Yeast extract reached 0.92% at 44 hours, and 0.96% at 32 hours, and 0.90% at 32 hours respectively, Also, that of Bios 2,000 reached 0.97% at 32 hours, which exhibited the highest, among the treatments. 5. The pH of control was 4.27 at 44 hours. that of CR starter medium was 4.33 at 40 hours and that of Yeast extract was 4.25 at 32 hours during the incubation in Streptococcus thermophilus. Besides, pH of Bios 2000 is lowest as 4.18 at 32 hours. 6. Lactic acid bacterial counts in control, CR starter medium, and Yeast extract during the incubation of Streptococcus thermophilus were $9.8{\times}10^{9}/ml$ at 44 hours,$9.5{\times}10^{8}/ml$ at 40 hours, and $9.6{\times}10^{8}/ml$ at 32 hours. And, the highest number was $2.0{\times}10^{9}/ml$ for Bios 2000 at 32 hours. 7. The acidity of control during the incubation of Lactobacillus casei reached 0.92% at 40 hours, and those of CR starter medium and Yeast extract were 0.95% at 40 hours, and 1.01% at 36 hours respectively. Also, Bios 2000 had the highest acidity as 0.94% at 32 hours. 8. The pH of control, CR starter medium and Yeast extract during the incubation Lactobacillus casei was 4.27 at 40 hours. 4.21 at 40 hours, and 4.15 at 36 hours respectively. Also, Bios 2000 showed the lowest pH, as 4.23, at 32 hours. 9. Lactic acid bacterial counts in control, CR starter medium and Yeast extract during the incubation of Lactobacillus casei were $9.4{\times}10^{7}/ml$ at 40 hours, $1.1{\times}10^{8}/ml$ at 40 hours, and $5.0{\times}10^{8}/ml$ at 36 hours respectively. And, the progress of 32 hours showed the highest number of lactic acid bacteria as$6.4{\times}10^{8}/ml$ in Bios 2000.

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Intensive Culture of the Pacific White Shrimp Litopenaeus vannamei under Limited Water Exchange I. Indoor Nursery Culture of Postlarvae (사육수 비교환 방식에 의한 흰다리새우의 고밀도 사육 I. 후기유생(postlarva)의 실내 중간육성)

  • Jang, In-Kwon;Kim, Jong-Sheek;Cho, Kook-Jin;Seo, Hyung-Chul;Cho, Yeong-Rok;Gopalakannan, Ayyaru;Kim, Bong-Lae
    • Journal of Aquaculture
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    • v.21 no.4
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    • pp.339-345
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    • 2008
  • Farming of the fleshy shrimp Fenneropenaeus chinensis which is a major cultured species in the west coast of South Korea, has been suffered :trom mass mortality due to disease epizootics including viruses. Since the Pacific white shrimp Litopenaeus vannamei was introduced to Korea in 2003, farming of this species has rapidly increased for years, occupying 62.5% of total cultured shrimp production in 2007. However the studies on L. vannamei culture methods for shrimp farming situations in Korea are very limited. Nursery culture of shrimp larvae has some advantages including increased survival, improved feed efficiencies, enhanced growth performance and reduced grow-out period. In this study, L. vannamei postlarvae (${PL_3}-{PL_{10}}$) with a density of $3,750-9,090/m^3$ were cultured in four raceways under limited water exchange condition for 35 days. Survival was the highest (93.6%) in tank stocked with $4,090/m^3$ and was the lowest in tank with $9,090/m^3$ (58.1 %). Mean body weight at harvest ranged from 0.071 to 0.108 g, and FCR was 0.59-0.70 in all tanks. Concentration of total ammonia nitrogen was increased up to 20 ppm on day 10 in all tanks and thereafter gradually decreased by the third week of culture. Nitrite-nitrogen was rapidly increased from the third week, representing bio-floc condition by developed nitrifying bacterial community. Of the present nursery system some modification of structure and consideration for commercial scale are needed in order to be implemented to shrimp farmers.

Immuno-stimulatory Activities of a High Molecular Weight Fraction from Cynanchum wilfordii Radix Obtained by Ultrafiltration (한외거르기(Ultrafiltration)에 의하여 분리된 백수오 고분자 분획물의 면역증진 활성)

  • Jang, Mi;Lim, Tae-Gyu;Hong, Hee-Do;Rhee, Young Kyoung;Kim, Kyung-Tack;Lee, Eunjung;Lee, Jeong Hoon;Lee, Yun Ji;Kim, Yeon Bok;Cho, Chang-Won
    • Korean Journal of Food Science and Technology
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    • v.48 no.3
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    • pp.268-274
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    • 2016
  • The purpose of this study was to investigate the immuno-stimulatory activity of the high-molecular-weight fraction (HMWF) of Cynanchum wilfordii (CW) extracts obtained by ultrafiltration in murine macrophage RAW 264.7 cells and to assess its immuno-stimulatory effect in mice. Ultrafiltration was performed with polyethersulfone membranes (30 kDa cutoff) in a cross-flow filtration system to obtain the HMWF of CW. The results showed that the HMWF increased the production of various cytokines such as tumor necrosis factor-${\alpha}$, interleukin-6, and nitric oxide in dose-ependent manners. In addition, HMWF treatment increased the relative spleen weight as well as splenocyte proliferation induced by concanavalin A or bacterial lipopolysaccharide in mice. Natural killer (NK) cell activity in the HMWF-treated group was significantly increased compared to that in the control group. These results suggest that the HMWF of CW can support the immune system through secretion of macrophage cytokines, thereby enhancing NK cell activity and murine splenocyte proliferation.

Isolation of Agarivorans sp. KC-1 and Characterization of Its Thermotolerant β-Agarase (한천분해세균 Agarivorans sp. KC-1의 분리 및 내열성 β-아가라제의 특성 규명)

  • Min, Kyung-Cheol;Lee, Chang-Eun;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.28 no.9
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    • pp.1056-1061
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    • 2018
  • This article reports an agar-degrading marine bacterium and characterizes its agarase. The agar-degrading marine bacterium, KC-1, was isolated from seawater on the shores of Sacheon, in Gyeongnam province, Korea, using Marine Broth 2216 agar medium. To identify the agar-degrading bacterium as Agarivorans sp. KC-1, phylogenetic analysis based on the 16S rRNA gene sequence was used. An extracellular agarase was prepared from a culture medium of Agarivorans sp. KC-1, and used for the characterization of enzyme. The relative activities at 20, 30, 40, 50, 60, and $70^{\circ}C$ were 65, 91, 96, 100, 77, and 35%, respectively. The relative activities at pH 5, 6, 7, and 8 were 93, 100, 87, and 82%, respectively. The extracellular agarase showed maximum activity (254 units/l) at pH 6.0 and $50^{\circ}C$ in 20 mM of Tris-HCl buffer. The agarase activity was maintained at 90% or more until 2 hr exposure at $20^{\circ}C$, $30^{\circ}C$ and $40^{\circ}C$, but it was found that the activity decreased sharply from $60^{\circ}C$. A zymogram analysis showed that Agarivorans sp. KC-1 produced 3 agar-degrading enzymes that had molecular weights of 130, 80, and 69 kDa. A thin layer chromatography analysis suggested that Agarivorans sp. KC-1 produced extracellular ${\beta}$-agarases as it hydrolyzed agarose to produce neoagarooligosaccharides, including neoagarohexaose (21.6%), neoagarotetraose (32.2%), and neoagarobiose (46.2%). These results suggest that Agarivorans sp. KC-1 and its thermotolerant ${\beta}$-agarase would be useful for the production of neoagarooligosaccharides that inhibit bacterial growth and delay starch degradation.

DEU-7 Derived from Ulmus macrocarpa Improved Immune Functions in Cyclophosphamide-treated Mice (면역억제 마우스 모델에서 왕느릅나무 유래 DEU-7의 면역기능 증강)

  • Kang, Kyung-Hwa;Go, Ji Su;Lee, Inhwan;Lee, Sang Ho;Lee, Sung Do;Kim, Deok Won;Lee, Jong-Hwan;Hwang, HyeJin;Hyun, Sook Kyung;KIM, Byoung Woo;Kim, Chul Min;Chung, Kyung Tae
    • Journal of Life Science
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    • v.25 no.10
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    • pp.1156-1163
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    • 2015
  • The present study investigated the immunomodulatory properties of four different medicinal plants in a cyclophosphamide-treated Balb/c mouse model. One of the four plants, Ulmus macrocarpa, showed partial resistance against immune suppression induced by cyclophosphamide. The bark of U. macrocarpa, commonly known as the Chinese elm, has been used as a pharmaceutical material in Korean traditional medicine to treat bacterial inflammation and induce wound healing. In this study, water extract of U. macrocarpa, named DEU-7, was used for its immunomodulating functional activity. DEU-7 increased the weight of the spleen and the number of splenocytes but did not significantly affect the liver, kidney, and thymus in vivo. A splenocyte viability assay confirmed that DEU-7 influenced ex vivo splenocyte survival. DEU-7 also increased the levels of cytokines, such as IL-2 and IL-4, and immunoglobulins, such as IgM, IgG, and IgA. These results indicated that DEU-7 is involved in the activation of T and B lymphocytes. In addition, DEU-7 was able to maintain the production of cytokines, such as TNF-α, IL-12, and IFN-γ, in the condition of cyclophosphamide-induced immune suppression, suggesting that DEU-7 activated innate immune cells, even under immune suppression. We concluded that DEU-7 aids immunological homeostasis, thereby preventing immune suppression, and aids both innate and adaptive immune response by maintaining the levels of various cytokines and immunoglobulins. Consequently, it is worth investigating the potential of DEU-7 as a supplemental source for immune-enhancing agents.

Characterization of B Cells of Lymph Nodes and Peripheral Blood in a Patient with Hyper IgM Syndrome (Hyper IgM Syndrome 환자에서 얻은 림프절 및 말초혈액 B세포의 특성)

  • Kim, Dong Soo;Shin, Kyuong Mi;Yang, Woo Ick;Shin, Jeon-Soo;Song, Chang Hwa;Jo, Eun Kyeong
    • Clinical and Experimental Pediatrics
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    • v.46 no.2
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    • pp.128-136
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    • 2003
  • Purpose : Hyper IgM syndrome(HIGM) is characterized by severe recurrent bacterial infections with decreased serum levels of IgG, IgA, and IgE but elevated IgM levels. Recently, it has been classified into three groups; HIGM1, HIGM2 and a rare form of HIGM. HIGM1 is a X-linked form of HIGM and has now been identified as a T-cell deficiency in which mutations occur in the gene that encodes the CD40 ligand molecule. HIGM2 is an autosomal recessive form of HIGM. Molecular studies have shown that the mutation of HIGM2 is in the gene that encodes activation-induced cytidine deaminase(AID). Recently, another rare form of X-linked HIGM syndrome associated with hypohydrotic ectodermal dysplasia has been identified. We encountered a patient with a varient form of HIGM2. To clarify the cause of this form of HIGM, we evaluated the peripheral B cells of this patient. Methods : The lymphocytes of the patient were prepared from peripheral blood. B cells were immortalized with the infection of EBV. Cell cycle analysis was done with the immortalized B cells of the patient. Peripheral mononuclear cells were stained with monoclonal anti-CD40L antibody. Total RNA was extracted from the peripheral mononuclear cells. After RT-PCR, direct sequencing for CD40L gene and HuAID gene were done. Immunostainings of a lymph node for CD3, CD23, CD40, Fas-L, bcl-2, BAX were done. Results : The peripheral B cells of this patient showed normal expression of CD40L molecule and normal sequencing of CD40L gene, and also normal sequencing of AID gene. Interestingly, the peripheral B cells of this patient showed a decreased population of G2/mitosis phase in cell cycles which recovered to normal with the stimulation of IL-4. Conclusion : We suspect that the cause of increased serum IgM in this patient may be from a decrease of G2/mitosis phase of the peripheral B cells, which may be from the decreased production or secretion of IL-4. Therefore, this may be a new form of HIGM.

MMP-1 and TIMP-1 production in MG-63 cells stimulated with Prevotella nigrescens Lipopolysaccharide (Prevotella nigrescens lipopolysaccharides로 자극된 MG63 세포에서 분비되는 기질금속단백질 MMP-1과 TIMP-1의 수준에 관한 연구)

  • Yang Won-Kyung;Kim Mi-Ri;Shon Won-Jun;Lee In-Bog;Cho Byeong-Hoon;Um Chung-Moon;Son Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.29 no.5
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    • pp.470-478
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    • 2004
  • The purpose of this study is to monitor the secretion of matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) produced by human osteosarcoma cell line (MG63) stimulated with Prevotella nigrescens lipopolysaccharides (LPS). and to compare the level of secretion before and after the treatment of calcium hydroxide on P. nigrescens LPS. LPS was extracted and purified from anaerobically cultured P. nigrescens. MG63 cells were stimulated by the LPS (0, 1, $10{\;}\mu\textrm{g}/ml$) or LPS($10{\;}\mu\textrm{g}/ml$) pretreated with 12.5 mg/ml of $Ca(OH)_2$ for 3 days. Total RNA was isolated from the cell. and real-time quantitative polymerase chain reaction (PCR) was performed for quantification of MMP-1 and TIMP-1. The results were as follows. 1. MMP-1 mRNA expression at 48 hr was highly increased by stimulation with P. nigrescens LPS. The increase was dose-dependent. 2. When stimulated with ($1{\;}\mu\textrm{g}/ml$ of LPS. TIMP-1 mRNA expression was highly increased at 24 hr and 48 hr. However. TIMP-1 expression was suppressed at higher concentration ($10{\;}\mu\textrm{g}/ml$). 3. When P. nigrescens LPS was pretreated with $Ca(OH)_2$. MMP-1 and TIMP-1 gene expression was downregulated. The results of this study suggest that transcriptional regulation of MMP-1 and TIMP-1 by P. nigrescens LPS could be one of the important mechanisms in bone resorption of periapical inflammation. The result of calcium hydroxide on MMP-1 and TIMP-1 gene expression suppression shows that calcium hydroxide detoxified bacterial LPS and thus should be used the medication of choice for intracanal dressings in root canal infected with black-pigmented bacteria.

Comparison of the Quantitative Values of C-14 and C-13 UBT to Reflect the Presence and Degree of Ongoing Helicobacter pylori Infection (Helicobacter pylori 감염 유무와 정도 반영에 대한 C-14와 C-13 요소호기검사 정량치 비교)

  • Lim, Seok-Tae;Kim, Dong-Wook;Jeong, Hwan-Jeong;Sohn, Myung-Hee
    • Nuclear Medicine and Molecular Imaging
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    • v.42 no.3
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    • pp.229-234
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    • 2008
  • Purpose: A urea breath test (UBT) using C-14 or C-13 has been developed for identifying Helicobacter (H) pylori infection on the basis of urease production with release of labeled $CO_2$. We investigated if the C-14 and C-13 UBT have the difference to reflect the presence and degree of H. pylori infection detected by gastro-duodenoscopic biopsies (CBx) in the same patients. Materials and methods: Thirty eight patients (M:F = 28:10, age $53.4{\pm}13.0$ yrs) with upper gastrointestinal symptoms such as indigestion, gastric fullness or pain consecutively underwent C-14 UBT, GBx and C-13 UBT within one week before medications. For the C-14 UBT, a single breath sample was collected at 10 minutes after ingestion of C-14 urea (37 KBq) capsule and counting was done in a liquid scintillation counter for 1 minute, and the results were classified as positive (${\ge}$ 200 dpm), intermediate (50-199 dpm) or negative (50 dpm). For the C-13 UBT, the results were classified as positive (${\ge}2.5\%_{\circ}$) or negative ($<2.5\%_{\circ}$). The results of GBx with Giemsa stain were graded 0 (normal) to 4 (diffuse) according to the distribution of H. pylori by the Wyatt method. We compared C-14 UBT and C-13 UBT results with GBx grade as a gold standard. Results: The prevalence of H. pylori infection by GBx with Giemsa stain was 25/38 (65.8%). In the assessment of the presence of H. pylori infection, the C-14 UBT global performance yielded sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy of 92.0%, 92.3%, 95.8%, 91.7% and 92.1%, respectively. However, the C-13 UBT had sensitivity, specificity, PPV, NPV and accuracy of 96.0%, 84.6%, 92.3%, 91.7% and 92.1%, respectively. The more significant correlation in C-14 than C-13 UBT (r=0.948 vs r=0.819, p <0.001) was found between the value of UBT and the grade of distribution of H. pylori infection. Conclusion: We conclude that the diagnostic performance between C-14 and C-13 UBT to detect H. pylori infection is not significantly different, but the value of C-14 UBT more significantly reflects the degree of bacterial distribution.

In vivo Antifungal Activity of Pyrrolnitrin Isolated from Burkholderia capacia EB215 with Antagonistic Activity Towards Colletotrichum Species (탄저병균에 대하여 길항작용을 보이는 Burkholderia cepacia EB215로부터 분리한 Pyrrolnitrin의 항균활성)

  • Park, Ji-Hyun;Choi, Gyung-Ja;Lee, Seon-Woo;Jang, Kyoung-Soo;Choi, Yong-Ho;Chung, Young-Ryun;Cho, Kwang-Yun;Kim, Jin-Cheol
    • The Korean Journal of Mycology
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    • v.32 no.1
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    • pp.31-38
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    • 2004
  • An endophytic bacterial strain EB215 that was isolated from cucumber (Cucumis sativus) roots displayed a potent in vivo antifungal activity against Colletotrichum species. The strain was identified as Burkholderia cepacia based on its physiological and biochemical characteristics, and 16S rDNA gene sequence. Optimal medium and incubation period for the production of antifungal substances by B. cepacia EB215 were nutrient broth (NB) and 3 days, respectively. An antifungal substance was isolated from the NB cultures of B. cepacia EB215 strain by centrifugation, n-hexane partitioning, silica gel column chromatography, preparative TLC, and in vitro bioassay. Its chemical structure was determined to be pyrrolnitrin by mass and NMR spectral analyses. Pyrrolnitrin showed potent disease control efficacy of more than 90% against pepper anthracnose (Colletotrichum coccodes), cucumber anthracnose (Colletotrichum orbiculare), rice blast (Magnaporthe grisea) and rice sheath blight (Corticium sasaki) even at a low concentration of $11.1\;{\mu}g/ml$. In addition, it effectively controlled the development of tomato gray mold (Botrytis cinerea) and wheat leaf rust (Puccinia recondita) at concentrations over $33.3\;{\mu}g/ml$. However, it had no antifungal activity against Phytophthora infestans on tomato plants. Further studies on the development of microbial fungicide using B. cepacia EB215 are in progress.

Production of a hypothetical polyene substance by activating a cryptic fungal PKS-NRPS hybrid gene in Monascus purpureus (홍국Monascus purpureus에서 진균 PKS-NRPS 하이브리드 유전자의 발현 유도를 통한 미지 polyene 화합물의 생성)

  • Suh, Jae-Won;Balakrishnan, Bijinu;Lim, Yoon Ji;Lee, Doh Won;Choi, Jeong Ju;Park, Si-Hyung;Kwon, Hyung-Jin
    • Journal of Applied Biological Chemistry
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    • v.61 no.1
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    • pp.83-91
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    • 2018
  • Advances in bacterial and fungal genome mining uncover a plethora of cryptic secondary metabolite biosynthetic gene clusters. Guided by the genome information, targeted transcriptional derepression could be employed to determine the product of a cryptic gene cluster and to explore its biological role. Monascus spp. are food grade filamentous fungi popular in eastern Asia and several genome data belong to them are now available. We achieved transcription activation of a cryptic fungal polyketide synthase-nonribosomal peptide synthase gene Mpfus1 in Monascus purpureus ${\Delta}MpPKS5$ by inserting Aspergillus gpdA promoter at the upstream of Mpfus1 through double crossover gene replacement. The gene cluster with Mpfus1 show a high similarity to those for the biosynthesis of conjugated polyene derivatives with 2-pyrrolidone ring and the mycotoxin fusarin is the representative member of this group. The ${\Delta}MpPKS5$ is incapable of producing azaphilone pigment, providing an excellent background to identify chromogenic and UV-absorbing compounds. Activation of Mpfus1 resulted in a yellow hue on mycelia and its methanol extract exhibit a maximum absorption at 365 nm. HPLC analysis of the organic extracts indicated the presence of a variety of yellow compounds in the extract. This implies that the product of MpFus1 is metabolically or chemically unstable. LC-MS analysis guided us to predict the MpFus1 product and to propose that the Mpfus1-containing gene cluster encode the biosynthesis of a desmethyl analogue of fusarin. This study showcases the genome mining in Monascus and the possibility to unveil new biological activities embedded in it.