Gene Expression Analysis of Phanerochaete chrysosporium During the Transition Time from Primary Growth to Secondary Metabolism

Mingfeng Jiang,Xiao Li,Liang Zhang,Hong Feng,Yizheng Zhang 한국미생물학회 2009 The journal of microbiology Vol.47 No.3

In order to identify the secondary metabolism-related genes of Phanerochaete chrysosporium growing under pure O2 and nitrogen-limited conditions, 2322 ESTs fragments originated from two suppression-subtractive libraries were analyzed using the cDNA microarray technique. Ten significantly upregulated and 22 significantly downregulated genes were identified in the 72 h cultured mycelia RNA samples (secondary metabolism). According to qPCR, 16 out of the 32 genes were expressed differently in secondary metabolism. Transcripts of secondary metabolism up-regulation genes exhibited homologies to aryl-alcohol dehydrogenase (SSh1554), ABC transporter gene (SSH624), chitinase (SSH963), heat shock protein (SSH1193), catalase (SSH317), cytochrome P450 (SSH331), glucosamine-6-phosphate isomerase (SSH611), and alkyl hydroperoxide reductase (SSH362) genes. Ninety-three genes could be classified by Eukaryotic Orthologous Groups (KOG). Among the genes assigned a function, gene expression patterns were different in both secondary metabolism and primary metabolism. In the group of “Cellular Processes and Signaling,” most of the genes were from the primary metabolism library. On the other hand, genes from the secondary metabolism library were found mainly in the “Information Storage” and “Processing and Poorly Characterized” groups. Based on the KOG functional assignments, six genes belong to the ubiquitin system, and all of them were from primary metabolism phase. The presence of the H2O2-relevant genes suggested that parts of the genes expressed in 72 h might be involved in the ligninolytic process during secondary metabolism of P. chrysosporium.

Systems biology and biotechnology of Streptomyces species for the production of secondary metabolites

Hwang, K.S.,Kim, H.U.,Charusanti, P.,Palsson, B.O.,Lee, S.Y. Pergamon Press ; Elsevier Science Ltd 2014 Biotechnology advances Vol.32 No.2

Streptomyces species continue to attract attention as a source of novel medicinal compounds. Despite a long history of studies on these microorganisms, they still have many biochemical mysteries to be elucidated. Investigations of novel secondary metabolites and their biosynthetic gene clusters have been more systematized with high-throughput techniques through inspections of correlations among components of the primary and secondary metabolisms at the genome scale. Moreover, up-to-date information on the genome of Streptomyces species with emphasis on their secondary metabolism has been collected in the form of databases and knowledgebases, providing predictive information and enabling one to explore experimentally unrecognized biological spaces of secondary metabolism. Herein, we review recent trends in the systems biology and biotechnology of Streptomyces species.

The putative C2H2 transcription factor RocA is a novel regulator of development and secondary metabolism in Aspergillus nidulans

Won Dong Chan,Kim Yong Jin,Kim Da Hye,Park Hee-Moon,맹필재 한국미생물학회 2020 The journal of microbiology Vol.58 No.7

Multiple transcriptional regulators play important roles in the coordination of developmental processes, including asexual and sexual development, and secondary metabolism in the filamentous fungus Aspergillus nidulans. In the present study, we characterized a novel putative C2H2-type transcription factor (TF), RocA, in relation to development and secondary metabolism. Deletion of rocA increased conidiation and caused defective sexual development. In contrast, the overexpression of rocA exerted opposite effects on both phenotypes. Additionally, nullifying rocA resulted in enhanced brlA expression and reduced nsdC expression, whereas its overexpression exerted the opposite effects. These results suggest that RocA functions as a negative regulator of asexual development by repressing the expression of brlA encoding a key asexual development activator, but as a positive regulator of sexual development by enhancing the expression of nsdC encoding a pivotal sexual development activator. Deletion of rocA increased the production of sterigmatocystin (ST), as well as the expression of its biosynthetic genes, aflR and stcU. Additionally, the expression of the biosynthetic genes for penicillin (PN), ipnA and acvA, and for terrequinone (TQ), tdiB and tdiE, was increased by rocA deletion. Thus, it appears that RocA functions as a negative transcriptional modulator of the secondary metabolic genes involved in ST, PN, and TQ biosynthesis. Taken together, we propose that RocA is a novel transcriptional regulator that may act either positively or negatively at multiple target genes necessary for asexual and sexual development and secondary metabolism.

Effect of romosozumab in premenopausal women with severe osteoporosis and anorexia nervosa

Kazuki Fujimoto,Narumi Maki,Daisuke Hashiba,Toshifumi Maeyama,Ryosuke Nakagawa,Hajime Arai,Seiji Ohtori 대한골다공증학회 2023 Osteoporosis and Sarcopenia Vol.9 No.4

Objectives: This study aims to investigate the effects of romosozumab on bone mineral density (BMD) and bone metabolism. Methods: In this retrospective case series, romosozumab was administered to 5 premenopausal female patients with osteoporosis and anorexia nervosa with fragility fractures. BMD and bone turnover marker changes were investigated at 6 months and 1 year after administering romosozumab. Results: BMD increased and high-turnover bone metabolism decreased 6 months and 1 year after administering romosozumab. Conclusions: Romosozumab is useful for treating osteoporosis in patients with anorexia nervosa.

Roles of Putative Sodium-Hydrogen Antiporter (SHA) Genes in S. coelicolor A3(2) Culture with pH Variation

( Yoon Jung Kim ),( Myung Hee Moon ),( Jae Sun Lee ),( Soon Kwang Hong ),( Yong Keun Chang ) 한국미생물 · 생명공학회 2011 Journal of microbiology and biotechnology Vol.21 No.9

Culture pH change has some important roles in signal transduction and secondary metabolism. We have already reported that acidic pH shock enhanced actinorhodin production in Streptomyces coelicolor. Among many potential governing factors on pH variation, the putative Na+/H+ antiporter (sha) genes in S. coelicolor have been investigated in this study to elucidate the association of the sha on pH variation and secondary metabolism. Through the transcriptional analysis and overexpression experiments on 8 sha genes, we observed that most of the sha expressions were promoted by pH shock, and in the opposite way the pH changes and actinorhodin production were enhanced by the overexpression of each sha. We also confirmed that sha8 especially has a main role in maintaining cell viability and pH homeostasis through Na+ extrusion, in salt effect experiment under the alkaline medium condition by deleting sha8. Moreover, this gene was observed to have a function of pH recovery after pH variation such as the pH shock, being able to cause the sporulation. However, actinorhodin production was not induced by the only pH recovery. The sha8 gene could confer on the host cell the ability to recover pH to the neutral level after pH variation like a pH drop. Sporulation was closely associated with this pH recovery caused by the action of sha8, whereas actinorhodin production was not due to such pH variation patterns alone.

Identification of the duplicated genes for <i>S</i>-adenosyl-<small>L</small>-methionine synthetase (<i>metK1-sp</i> and <i>metK2-sp</i>) in <i>Streptomyces peucetius</i> var. <i>caesius</i> ATCC 27952

Oh, T.-J.,Niraula, N.P.,Liou, K.,Sohng, J.K. Blackwell Publishing Ltd 2010 Journal of applied microbiology Vol.109 No.2

<P>Abstract</P><P>Aims: </P><P>To characterize the function of both <I>metK1-sp</I> (<I>sp1190</I>) and <I>metK2-sp</I> (<I>sp1566</I>) <I>in vitro</I> and <I>in vivo</I>, and to study the regulation of doxorubicin production by overexpressing the <I>metK</I>.</P><P>Methods and Results: </P><P>We cloned two <I>orfs</I> into pET32a(+) respectively, and the formation of <I>S</I>-Adenosyl-<SMALL>L</SMALL>-methionine was clearly observed in the <I>in vitro</I> enzyme assays as functional MetKs. Reverse transcriptase polymerase chain reaction (PCR) analysis indicated that the transcripts for the <I>metK1-sp</I> were repressed as <I>Streptomyces</I> cells entered the decline phase, whereas that of the <I>metK2-sp</I> was induced, suggesting that these MetK proteins may be important for the growth and the regulation of secondary metabolites during the stationary growth phase, whether considered together or separately. Furthermore, we found that the introduction of high-copy-number plasmids containing the <I>metK1-sp</I> and <I>metK2-sp</I> resulted in 2·1- and 1·4-fold greater levels of doxorubicin production than the control transformants containing only the vector, respectively. We also attempted to disrupt the <I>metK-sp</I> and found that doxorubicin production from the <I>metK1-sp</I>-deleted mutant (<I>Streptomyces peucetius/pNN1</I>) was reduced when compared to the parent strain (<I>S. peucetius</I> var. <I>caesius</I> ATCC 27952).</P><P>Conclusions: </P><P>The results of this study indicated that two <I>metK</I> are differentially expressed during cell growth, and that the expressions of the two <I>metK</I> genes are differentially regulated under the same conditions.</P><P>Significance and Impact of the Study: </P><P><I>Streptomyces peucetius</I> var. <I>caesius</I> contains two genes, <I>metK1-sp</I> and <I>metK2-sp</I>, which encode functional <I>S</I>-adenosyl-<SMALL>L</SMALL>-methionine synthetase (MetK). The degree of homology (90% identity) found between the two genes shows that <I>metK1-sp</I> and <I>metK2-sp</I> are duplicated genes. Although there is currently no evidence for the relationship of the duplicated <I>metK</I> genes involved in the regulation of doxorubicin production, <I>metK1-sp</I> and <I>metK2-sp</I> may play a role in controlling the stimulation of antibiotic production during secondary metabolism.</P>

대사유도물질 처리에 의한 발아녹두의 아이소플라본 생합성 양상

이지현,정일민,박세준,김욱한,김소연,김진애,정우식 韓國作物學會 2004 Korean journal of crop science Vol.49 No.6

발아 녹두에 세 가지 스트레스 관련 화합물 salicylic acid, methyl jasmonic acid, acetyl salicylic acid를 처리하여 isoflavone의 생합성양상을 관찰한 결과를 요약하면 다음과 같다. 1. 숙주나물의 자엽에서는 isoflavone총량이 건조중 1g당 832.5ug 인 무처리구와 비교하여 10mM salicylic acid를 처리한 경우 169~% , 12mM acetyl salicylic acid로 처리한 경우 165~% 의 isoflavone 총량이 증가한 반면 0.5~% methyl jasmonic acid를 처리한 경우는 오히려 무처리구보다 47~% 수준으로 감소하였다. 2. 숙주나물의 자엽하부(hypocotyl and root)의 isoflavone 생성량에서는 1g당 284.8ug 이 생성된 무처리구와 비교하여 세 가지 처리 모두에서 유의성이 있는 차이를 보였다. 10mM salicylic acid 처리구의 경우 419~% , 12mM acetyl salicylic acid 처리구의 경우 401~% 의 isoflavone 총량의 증가를 보였고, 0.5~% methyl jasmonic acid처리구의 경우에는 121~% 증가하였다. 3.숙주나물의 자엽부위와 자엽하부에서 검출된 isoflavone의 합을 각 처리별 isoflavone생산총량으로 하여 무처리구의 건조중 1g당 1117.3ug 을 기준으로 비교하여보면 건조중 1g당 10mM salicylic acid 처리구에서는 2601.02ug 으로 233~% 증가하였고, 12mM acetyl salicylic acid 2514.4ug 으로 225~% 증가한 반면, 0.5~% methyl jasmonic acid 처리구에서는 738.8ug 으로 66~% 수준으로 감소하였다. 4. 숙주나물 자엽부위의 경우 무처리구와 비교하여 증가를 보였던 10mM salicylic acid처리구와 12mM acetyl salicylic acid 처리구에서는 malonyldaidzine과 malonylglycitin이 증가가 두드러지게 나타났다. 5. 숙주나물 자엽하부의 경우 무처리구와 비교하여 증가를 보였던 10mM salicylic acid 처리구와 12mM acetyl salicylic acid 처리구에서는 malonylglycitin의 증가가 두드러지게 나타났다 We have studied physiological responses of mung bean sprout to the treatment of elicitors. Chemicals such as salicylic acid and methyl jasmonic acid are not only the intermediates found in plant defense system but also could affect plant secondary metabolism. We found that mild treatment of salicylic acid and acetyl salicylic acid (aspirin) increase isoflavone production dramatically in mung bean sprout which has very low level of isoflavones compared with soybean sprout. The isoflavone content in salicylic acid treated- and acetyl salicylic acid treated-mung bean sprout was about 2.3 and 2.2 times higher than that of control, respectively. However, the increasing patterns of isoflavone in cotyledon and hypocotyl and root were not identical. The major increase among isoflavone fractions in cotyledon was led by the increase in malonylglycitin and malonyldaidzin level. Whereas, the increase in hypocotyl and root was led by malonyldaidzin. Methyl jasmonic acid did not show statistically significant increase in mung bean sprout. With this result, we were able to propose the non-transgenic method, which can control the isoflavone production in germinating mung bean

‘홍로’ 사과 과피 및 과육의 단백질체 비교 분석

정정성(Jung-Sung Chung),최성환(Sung Hwan Choi),김재호(Jae Ho Kim),심수용(Soo Yong Shim),모크 아드난(Moch R. Adnan),장은실(Eun-Sil Chang),손영걸(Young-Geol Sohn),김윤희(Yun-Hee Kim),김진국(Jin Gook Kim),이증주(Jeung Joo Lee) 한국원예학회 2021 원예과학기술지 Vol.39 No.2

국내 육종 사과 품종인 ‘홍로’의 과피와 과육에서 발현되는 단백질체를 추출하여 이차원 전기영동으로 분리하였다. 그 결과 대부분의 단백질들은 과피와 과육에서 유사한 정도로 발현되었으나, 30개의 단백질 spot들은 과피 또는 과육에서의 발현 정도가 2배 이상 차이가 났다. 이들 중 25개는 과피에서, 5개는 과육에서 더 높게 발현되었다. 이들 단백질들을 MALDI-TOF/TOF로 분석한 결과, 과피에서 더 높게 발현된 단백질 spot들 중 18개는 각각의 기능을 갖는 단백질로 동정 되었고, 나머지 7개는 기능이 밝혀지지 않은 것으로 동정되었거나 미동정 되었다. 과육에서 더 높게 발현된 단백질 spot들의 경우 2개는 각각의 기능을 갖는 단백질로 동정 되었고, 나머지 3개는 미동정 되었다. 과피에서 더 높게 발현된 단백질은 에틸렌의 생합성과 관련된 1-aminocyclopropane-1-carboxylate oxidase homolog(spot 21번), 이차대사산물의 생합성과 관련된 acetyl-CoA acetyltransferase, cytosolic 1(spot 148), acetyl-CoA acetyltransferase, cytosolic 1-like(spot 316) 및 8-hydroxygeraniol dehydrogenase-like(spot 348), 광합성과 관련된 oxygen-evolving enhancer protein 1, chloroplastic(spot 210) 및 ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit(spot 273), 생체방어 및 스트레스 반응과 관련된 (+)-neomenthol dehydrogenase-like(spot 216 및 219), formate dehydrogenase, mitochondrial(spot 305), glutathione S-transferase-like(spot 483 및 484) 및 acidic endochitinase SE 2-like, partial(spot 391 및 392), 항산화효소인 L-ascorbate peroxidase 2(spot 221), 알레르기 관련 단백질인 MLP-like protein 34(spot 406), major allergen Mal d 1(spot 451 및 452) 및 major allergen Mal d 1.06A01(spot 457)이었다. 한편 과육에서 더 높게 발현된 단백질은 protein folding과 관련된 chaperonin 60 subunit alpha 1, chloroplastic(spot 8) 및 탄수화물 대사와 관련된 NADP-dependent D-sorbitol-6-phosphate dehydrogenase(spot 380)이었다. 이상의 연구 결과들은 사과의 육종, 저장, 품질, 환경에 대한 내성 등과 관련된 광범위한 연구에 유용하게 이용될 수 있을 것으로 사료되었다. The proteome expressed in the peel and flesh of ‘Hongro’, a domestic breeding apple cultivar, was extracted and separated by two-dimensional electrophoresis. Most of the proteins were expressed at similar levels in the peel and flesh, but 30 protein spots showed a difference of more than two times higher in expression level in the peel and flesh. Of these, 25 were higher in the peel and 5 were higher in the flesh. By analyzing these proteins with MALDI-TOF/TOF, 18 of the protein spots that were higher in the peel were identified as proteins with their respective functions, and the remaining 7 were identified as having unknown function or were unidentified. In the case of the protein spots expressed higher in the flesh, two were identified as proteins having respective functions, and the other three were unidentified. Proteins more highly expressed in the peel were 1-aminocyclopropane-1-carboxylate oxidase homolog (spot 21) related to the biosynthesis of ethylene; acetyl-CoA acetyltransferase (spot 148), cytosolic 1, acetyl-CoA acetyltransferase, cytosolic 1-like (spot 316) and 8-hydroxygeraniol dehydrogenase-like (spot 348) related to the biosynthesis of secondary metabolites; oxygen-evolving enhancer protein 1, chloroplastic (spot 210), and ribulose-1, 5-bisphosphate carboxylase/oxygenase large subunit (spot 273) related to the photosynthesis;(+)-neomenthol dehydrogenase-like (spot 216 and 219), formate dehydrogenase, mitochondrial (spot 305), glutathione S-transferase-like (spot 483 and 484), and acidic endochitinase SE 2-like, partial (spot 391 and 392) related to the biological defense and stress response; and antioxidant enzyme L-ascorbate peroxidase 2 (spot 221), MLP-like protein 34 (spot 406), major allergen Mal d 1 (spot 451 and 452), and major allergen Mal d 1.06A01 (spot 457) related to allergies. On the other hand, proteins that were more highly expressed in the flesh were chaperonin 60 subunit alpha 1, chloroplastic (spot 8) related to protein folding, and NADP-dependent D-sorbitol-6-phosphate dehydrogenase (spot 380) related to carbohydrate metabolism. These results will be useful in a wide range of studies related to apple breeding, storage, quality, and environmental tolerance.

Response of Carbon and Nitrogen Metabolism and Secondary Metabolites to Drought Stress and Salt Stress in Plants

Gaochang Cui,Yu Zhang,Wenjin Zhang,Duoyong Lang,Xiaojia Zhang,Zhixian Li,Xinhui Zhang 한국식물학회 2019 Journal of Plant Biology Vol.62 No.6

Carbon and nitrogen metabolism provide the mainenergy and basic nutrients for plants. However, environmentalstress seriously affects carbon and nitrogen metabolism and thushinders plant growth, especially drought stress and salt stress. Hence, numerous studies have been conducted to investigate theresponse of carbon and nitrogen metabolism to drought stressand salt stress by photosynthesis, sucrose and starch metabolism,nitrogen uptake and amino acids. Previous researchers alsostudied the response of secondary metabolism under bothstresses on account of secondary metabolism may conferprotection against environmental stresses. Our review highlightsthe diverse responses of carbon and nitrogen metabolism todrought stress and salt stress and the content changes of threesecondary metabolites in plants under stresses.

Genomics Reveals Traces of Fungal Phenylpropanoid-flavonoid Metabolic Pathway in the Filamentous Fungus Aspergillus oryzae

Juvvadi Praveen Rao,Seshime Yasuyo,Kitamoto Katsuhiko The Microbiological Society of Korea 2005 The journal of microbiology Vol.43 No.6

Fungal secondary metabolites constitute a wide variety of compounds which either playa vital role in agricultural, pharmaceutical and industrial contexts, or have devastating effects on agriculture, animal and human affairs by virtue of their toxigenicity. Owing to their beneficial and deleterious characteristics, these complex compounds and the genes responsible for their synthesis have been the subjects of extensive investigation by microbiologists and pharmacologists. A majority of the fungal secondary metabolic genes are classified as type I polyketide synthases (PKS) which are often clustered with other secondary metabolism related genes. In this review we discuss on the significance of our recent discovery of chalcone synthase (CHS) genes belonging to the type III PKS superfamily in an industrially important fungus, Aspergillus oryzae. CHS genes are known to playa vital role in the biosynthesis of flavonoids in plants. A comparative genome analyses revealed the unique character of A. oryzae with four CHS-like genes (csyA, csyB, csyC and csyD) amongst other Aspergilli (Aspergillus nidulans and Aspergillus fumigatus) which contained none of the CHS-like genes. Some other fungi such as Neurospora crassa, Fusarium graminearum, Magnaporthe grisea, Podospora anserina and Phanerochaete chrysosporium also contained putative type III PKSs, with a phylogenic distinction from bacteria and plants. The enzymatically active nature of these newly discovered homologues is expected owing to the conservation in the catalytic residues across the different species of plants and fungi, and also by the fact that a majority of these genes (csyA, csyB and csyD) were expressed in A. oryzae. While this finding brings filamentous fungi closer to plants and bacteria which until recently were the only ones considered to possess the type III PKSs, the presence of putative genes encoding other principal enzymes involved in the phenylpropanoid and flavonoid biosynthesis (viz., phenylalanine ammonia-lyase, cinnamic acid hydroxylase and p-coumarate CoA ligase) in the A. oryzae genome undoubtedly prove the extent of its metabolic diversity. Since many of these genes have not been identified earlier, knowledge on their corresponding products or activities remain undeciphered. In future, it is anticipated that these enzymes may be reasonable targets for metabolic engineering in fungi to produce agriculturally and nutritionally important metabolites.