• 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.

• Journal of Korea Technical Association of The Pulp and Paper Industry
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• v.46 no.1
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• pp.18-28
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• 2014

Renewable Portfolio Standards(RPS) is a regulation that requires a renewable energy generated from eco-friendly energy sources such as biomass, wind, solar, and geothermal. The RPS mechanism generally is an obligatory policy that places on electricity supply companies to produce a designated fraction of their electricity from renewable energies. The domestic companies to supply electricity largely rely on wood pellets in order to implement the RPS in spite of undesirable situation of lack of wood resources in Korea. This means that the electricity supply companies in Korea must explore new biomass as an alternative to wood. Palm kernel shell (PKS) and empty fruit bunch (EFB) as oil palm wastes can be used as raw materials used for making pellets after their thermochemical treatment like torrefaction. Torrefaction is a pretreatment process which serves to improve the properties including heating value and energy densification of these oil palm wastes through a mild pyrolysis at temperature typically ranging between 200 and $300^{\circ}C$ in the absence of oxygen under atmospheric pressure. Torrefaction of oil palms wastes at above $200^{\circ}C$ contributed to the increase of fixed carbon with the decrease of volatile matters, leading to the improvement of their calorific values over 20.9 MJ/kg (=5,000 kcal/kg) up to 25.1 MJ/kg (=6,000 kcal/kg). In particular, EFB sensitively responded to torrefaction because of its physical properties like fiber bundles, compared to PKS and hardwood chips. In conclusion, torrefaction treatment of PKS and EFB can greatly contribute to the implement of RPS of the electricity supply companies in Korea through the increased co-firing biomass with coal.

• Journal of Korea Technical Association of The Pulp and Paper Industry
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• v.47 no.1
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• pp.24-34
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• 2015

Domestic companies supplying electricity must increase obligatory duty to use renewable energy annually. If not met with obligatory allotment, the electricity-supply companies must pay RPS (Renewable Portfolio Standards) penalty. Although the power plants using a pulverizing coal firing boiler could co-fire up to around 3 per cent with wood pellets mixed in with coal feedstock without any major equipment revamps, they recorded only about 60 per cent fulfillment of RPS. Consequently, USD 46 million of RPS penalty was imposed on the six power supplying subsidiaries of GENCOs in 2014. One of the solutions to reduce the RPS penalty is that the power supply companies adopt the co-firing of torrefied lignocellulosic biomass in coal plants, which may contribute to the use of over 30 per cent of torrefied biomass mixed with bituminous coals. Extra binder was required to form pellets using torrefied biomass such as wood chips, PKS (Palm Kernel Shell) and EFB (Empty Fruit Bunch). Instead of corn starch, 30, 50 and 70 per cent of Larix saw dusts were respectively added to the torrefied feedstocks such as Pinus densiflora chips, PKS and EFB. The addition of saw dusts led to the decrease of the calorific values of the pellets but the forming ability of the pelletizer was exceedingly improved. Another advantage from the addition of saw dusts stemmed from the reduction of ash contents of the pellets. Finally, it was confirmed that torrefied oil palm biomass such as PKS and EFB could be valuable feedstocks in making pellets through improved binding ability.

• Particle and aerosol research
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• v.17 no.4
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• pp.125-132
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• 2021

Fresh PM_2.5 smokes emitted from combustion of four biomass materials (pellet, palm fruit fiber (PFF), PKS, and sawdust) in a laboratory-controlled environment were characterized using an attenuated total reflectance-fourier transform infrared (ATR-FTIR) technique. In smoke samples emitted from combustion of pellets, PFF and PKS, which is being used as boiler fuels for greenhouses in rural areas, the organic carbon/elemental carbon (OC/EC) ratios in PM_2.5 were very high (14.0-35.5), whereas in sawdust smoke samples they were significantly low (<4.0) due to the combustion method close to flaming combustion. ATR-FTIR analysis showed that OH(3400-3250 cm^-1), CH₃(2958-2840 cm^-1), CH₂(2910 cm^-1 and 2850 cm^-1), ketone(1726-1697 cm^-1), C=C(1607-1606 cm^-1 and 1515-1514 cm^-1), lignin (1463-1462 cm^-1 and 1430-1428 cm^-1) and -NO₂(1360-1370 cm^-1) peaks were identified in all biomass burning (BB) smoke samples. However, additional peaks appeared depending on the type of biomass. Among the four types of biomass materials, an additional peak of the methylene group CH₃(2872-2870 cm^-1) appeared only in PFF and PKS smoke samples, and a peak of C=O(1685 cm^-1) was also confirmed. And in the case of PKS smoke samples, a peak of aromatic C=C(1593 cm^-1 and 1476 cm^-1) that did not appear in other BB samples was also observed. This indicates that the molecular structure of organic compounds emitted during BB differs depending on the type of biomass materials. The results of this study are expected to provide valuable information to more specifically reveal the effect of BB on PM_2.5 collected in the atmospheric environment.

• The Journal of the Convergence on Culture Technology
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• v.8 no.6
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• pp.921-926
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• 2022

Sugar solutions (5%, 10%, 15% and 20%) were tested by seven sensors of Astree E-Tongue for selecting a sensor for sweetness. NMS sensor was chosen as a sensor for sweetness among two sensors (PKS and NMS sensors selected in first stage) by considering precision, linearity and accuracy. Sugar, fructose, glucose and xylitol (5%, 10% and 15%) were tested by E-tongue. The principal component analysis (PCA) result by E-Tongue with seven sensors at 5% concentration level of four sweetners was not satisfactory (Discrimination index was -0.1). On the other hand, the relative NMS sensor response values were derived as 1.08 (fructose), 0.99 (glucose) and 1.00 (xylitol) comparing to sugar. Only the E-Tongue relative glucose response 0.99 was different from 0.5~0.75 of the relative sweetness range reported as the human sensory test results. Considering the excellent precision (%RSD, 1.53~3.64%) of E-Tongue using NMS single sensor for three types of sweeteners compared to sugar in the concentration range of 5% to 15%, replacing sensory test of sweetened beverages by E-Tongue might be possible for new product development and quality control.

• Mycobiology
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• v.51 no.1
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• pp.36-48
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• 2023

Xanthoria elegans is a lichen symbiosis, that inhabits extreme environments and can absorb UV-B. We reported the de novo sequencing and assembly of X. elegans genome. The whole genome was approximately 44.63 Mb, with a GC content of 40.69%. Genome assembly generated 207 scaffolds with an N50 length of 563,100 bp, N90 length of 122,672 bp. The genome comprised 9,581 genes, some encoded enzymes involved in the secondary metabolism such as terpene, polyketides. To further understand the UV-B absorbing and adaptability to extreme environments mechanisms of X. elegans, we searched the secondary metabolites genes and gene-cluster from the genome using genome-mining and bioinformatics analysis. The results revealed that 7 NR-PKSs, 12 HR-PKSs and 2 hybrid PKS-PKSs from X. elegans were isolated, they belong to Type I PKS (T1PKS) according to the domain architecture; phylogenetic analysis and BGCs comparison linked the putative products to two NR-PKSs and three HR-PKSs, the putative products of two NR-PKSs were emodin xanthrone (most likely parietin) and mycophelonic acid, the putative products of three HR-PKSs were soppilines, (+)-asperlin and macrolactone brefeldin A, respectively. 5 PKSs from X. elegans build a correlation between the SMs carbon skeleton and PKS genes based on the domain architecture, phylogenetic and BGC comparison. Although the function of 16 PKSs remains unclear, the findings emphasize that the genes from X. elegans represent an unexploited source of novel polyketide and utilization of lichen gene resources.

• Journal of Microbiology
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• v.44 no.6
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• pp.649-654
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• 2006

A standard type II polyketide synthase (PKS) gene cluster was isolated while attempting to clone the biosynthetic gene for lipstatin from Streptomyces toxytricini NRRL 15,443. This result was observed using a Southern blot of a PstI-digested S. toxytricini chromosomal DNA library with a 444 bp amplified probe of a ketosynthase (KS) gene fragment. Four open reading frames [thioesterase (TE), $\beta$-ketoacyl systhase (KAS), chain length factor (CLF), and acyl carrier protein (ACP)], were identified through the nucleotide sequence determination and analysis of a 4.5 kb cloned DNA fragment. In order to confirm the involvement of a cloned gene in lipstatin biosynthesis, a gene disruption experiment for the KS gene was performed. However, the resulting gene disruptant did not show any significant difference in lipstatin production when compared to wild-type S. toxytricini. This result suggests that lipstatin may not be synthesized by a type II PKS.

• Journal of Microbiology and Biotechnology
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• v.16 no.1
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• pp.153-157
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• 2006

A soil metagenomic library was constructed using an E. coli-fosmid cloning system with environmental DNAs extracted from Kwangreung forest topsoil. We targeted the genes involved in the biosynthesis of bacterial polyketides. Initially, a total of 36 clone pools (10,800 clones) were explored by the PCR-based method using the metagenomic DNAs from each pool and a degenerate primer set, which has been designed based on the highly conserved regions among ketoacyl synthase (KS) domains in actinomycete type I polyketide synthases (PKS Is). Six clone pools were tentatively selected as positive and further examined through a hybridization-based method for selecting a fosmid clone containing PKS I genes. Colony hybridization was performed against fosmid clones from the 6 positive pools, and finally 4 clones were picked out and confirmed to contain the conserved DNA fragment of KS domains. In this study, we present a simple and feasible sorting method for a desired clone from metagenomic libraries.

• Molecules and Cells
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• v.38 no.12
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• pp.1105-1110
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• 2015

Phleichrome, a pigment produced by the phytopathogenic fungus Cladosporium phlei, is a fungal perylenequinone whose photodynamic activity has been studied intensively. To determine the biological function of phleichrome and to engineer a strain with enhanced production of phleichrome, we identified the gene responsible for the synthesis of phleichrome. Structural comparison of phleichrome with other fungal perylenequinones suggested that phleichrome is synthesized via polyketide pathway. We recently identified four different polyketide synthase (PKS) genes encompassing three major clades of fungal PKSs that differ with respect to reducing conditions for the polyketide product. Based on in silico analysis of cloned genes, we hypothesized that the non-reducing PKS gene, Cppks1, is involved in phleichrome biosynthesis. Increased accumulation of Cppks1 transcript was observed in response to supplementation with the application of synthetic inducer cyclo-(${_L}-Pro-{_L}-Phe$). In addition, heterologous expression of the Cppks1 gene in Cryphonectria parasitica resulted in the production of phleichrome. These results provide convincing evidence that the Cppks1 gene is responsible for the biosynthesis of phleichrome.

• KSBB Journal
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• v.22 no.5
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• pp.356-361
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• 2007

Cyanobacteria are a very old group of prokaryotic organisms that produce very diverse secondary metabolites, especially non-ribosomal peptide and polyketide structures. Although some cyanobacteria produce lethal toxins such as microcystins and anatoxins, some may be useful either for development into commercial drugs or as biochemical tools. Detection of unknown secondary metabolites was carried in the present study by a screening of 98 cyanobacterial strains from Cyanobiotech GmbH in order to establish a screening process, isolate pure substances and determine their bioactivities. A degenerated polymerase chain reaction technique as molecular approaches has been used for general screening of NRPS gene and PKS gene in cyanobacteria. A putative PKS gene was detected by DKF/DKR primer in 38 strains (38.8%) and PCR amplicons resulted from a presence of NRPS gene were showed by MTF2/MTR2 primer in 30 strains (30.6%), respectively. A screening of interesting strains was performed by comparing PCR screening results with HPLC analyses of extracts. HPLC analysis for a detection of natural products was performed in extracts from biomass. 5 strains were screened for further scale-up processing. 7 pure substances were isolated from the scale-up cultures and tested for bioactivities under consideration to purity, amount and molecular weight of substances. One substance isolated from CBT 635 showed cytotoxic activity. This substance may be regarded as Microcystin LR.