Journal of Microbiology

Journal Articles

Upgrading Isoquercitrin Concentration via Submerge Fermentation of Mulberry Fruit Extract with Edible Probiotics to Suppress Gene Targets for Controlling Kidney Cancer and Inflammation: Md Rezaul Karim, Safia Iqbal, Shahnawaz Mohammad, Jong-Hoon Kim, Li Ling, Changbao Chen, Abdus Samad, Md Anwarul Haque, Deok-Chun Yang, Yeon Ju Kim, Dong Uk Yang; J. Microbiol. 2024;62(10):919-927. Published online October 8, 2024; DOI: https://doi.org/10.1007/s12275-024-00163-8

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In recent years, kidney cancer has become one of the most serious medical issues. Kidney cancer is treated with a variety of active compounds that trigger genes that cause cancer. We identified in our earlier research that isoquercitrin (IQ) can activate PIK3CA, IGF1R, and PTGS2. However, it has a very low bioavailability because of its lower solubility in water. So, we utilized sub-merge fermentation technology with two well-known probiotics, Lactobacillus acidophilus and Bacillus subtilis, as a microbial source and mulberry fruit extract as a substrate, which has a high IQ level to improve IQ yield. Furthermore, we compared the total phenolic, flavonoid, and antioxidant contents of fermented and non-fermented samples, and we found that the fermented samples had greater levels than non-fermented sample. In addition, the high-performance liquid chromatography (HPLC) results showed that the fermented mulberry fruit extract from B. subtilis and L. acidophilus showed higher IQ values (190.73 ± 0.004 μg/ml and 220.54 ± 0.007 μg/ml, respectively), compared to the non-fermented samples, which had IQ values (80.12 ± 0.002 μg/ml). Additionally, at 62.5 µg/ml doses of each sample, a normal kidney cell line (HEK 293) showed higher cell viability for fermented and non-fermented samples. Conversely, at the same doses, the fermented samples of L. acidophilus and B. subtilis in a kidney cancer cell line (A498) showed an inhibition of cell growth around 36% and 31%, respectively. Finally, we performed RT and qRT PCR assay, and we found a significant reduction in the expression of the PTGS2, PIK3CA, and IGF1R genes. We therefore can conclude that the fermented samples have a higher concentration of isoquercitrin, and also can inhibit the expression of the genes PTGS2, PIK3CA, and IGF1R, which in turn regulates kidney cancer and inflammation.

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Recent research on the bioactivity of polyphenols derived from edible fungi and their potential in chronic disease prevention
Wenbin Yu, Yufei Zhang, Yi Lu, Zhiwei Ouyang, Jiahua Peng, Yayi Tu, Bin He
Journal of Functional Foods.2025; 124: 106627. CrossRef
Protective roles of genistein and icaritin in skin barrier integrity and hydration in an atopic dermatitis model
Shahnawaz Mohammad, Anjali Kariyarath Valappil, Md. Rezaul Karim, Safia Iqbal, Deok Chun Yang, Changbao Chen, Li Ling, Dong Uk Yang
European Journal of Integrative Medicine.2025; 76: 102483. CrossRef
Isoquercitrin Improves Insulin Resistance by Inhibiting PTP1B-Regulated IRS/PI3K/AKT Signaling Pathway
Si-yu Liu, Lu-jing Yu, Sheng-nan Zhang, Jia-kui Yue, Guo-jun Jiang, Si-hua Lu, Ying-jia Li, Jun-yu Meng, Gui-hong Huang
Chinese Journal of Integrative Medicine.2025;[Epub] CrossRef

Enhanced Poly-γ-Glutamic Acid Production by a Newly Isolated Bacillus halotolerans F29: Xiaorong Sun, Yaoyu Cai, Dexin Wang; J. Microbiol. 2024;62(8):695-707. Published online August 20, 2024; DOI: https://doi.org/10.1007/s12275-024-00153-w

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Abstract PDF

Poly-γ-glutamic acid (γ-PGA) is a promising biopolymer for various applications. In this study, we isolated a novel γ-PGA-producing strain, Bacillus halotolerans F29. The one-factor-at-a-time method was used to investigate the influence of carbon sources, nitrogen sources, and culture parameters on γ-PGA production. The optimal carbon and nitrogen sources were sucrose and (NH₄)₂SO₄, respectively. The optimal culture conditions for γ-PGA production were determined to be 37 °C and a pH of 5.5. Response surface methodology was used to determine the optimum medium components: 77.6 g/L sucrose, 43.0 g/L monosodium glutamate, and 2.2 g/L K₂HPO₄. The γ-PGA titer increased significantly from 8.5 ± 0.3 g/L to 20.7 ± 0.7 g/L when strain F29 was cultivated in the optimized medium. Furthermore, the γ-PGA titer reached 50.9 ± 1.5 g/L with a productivity of 1.33 g/L/h and a yield of 2.23 g of γ-PGA/g of L-glutamic acid with the optimized medium in fed-batch fermentation. The maximum γ-PGA titer reached 45.3 ± 1.1 g/L, with a productivity of 1.06 g/L/h when molasses was used as a carbon source. It should be noted that the γ-PGA yield in this study was the highest of all reported studies, indicating great potential for the industrial production of γ-PGA.

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Investigation of glutamic acid production capacity of Stenotrophomonas sp. strain CG2 isolated from soil
Cihat Guner, Ertan Ermis, Kubra Ozkan Guner
Biocatalysis and Agricultural Biotechnology.2025; 67: 103665. CrossRef
Metabolic engineering of microorganisms for tailor-made biopolymer production: A review
Mădălina Lorena Medeleanu, Lavinia-Florina Călinoiu, Gheorghe-Adrian Martău, Dan-Cristian Vodnar
International Journal of Biological Macromolecules.2025; 330: 147922. CrossRef
Poly-γ-glutamic acid production from untreated sugarcane molasses by non-sterilized repeated-batch fermentation with Bacillus subtilis GLS-8
Yu Lin, Lin Shu, Huizhen Chen, Xiaoqun Duan, Wei Zeng
Chemical Engineering Journal Advances.2025; 24: 100900. CrossRef
Transcriptomics-guided rational engineering in Bacillus licheniformis for enhancing poly-γ-glutamic acid biosynthesis using untreated molasses
Rui Han, Qian Zhong, Yifan Yan, Juan Wang, Yifan Zhu, Sha Li, Peng Lei, Rui Wang, Yibin Qiu, Zhengshan Luo, Hong Xu
International Journal of Biological Macromolecules.2024; 282: 137514. CrossRef

Delineating the Acquired Genetic Diversity and Multidrug Resistance in Alcaligenes from Poultry Farms and Nearby Soil: Abhilash Bhattacharjee, Anil Kumar Singh; J. Microbiol. 2024;62(7):511-523. Published online June 21, 2024; DOI: https://doi.org/10.1007/s12275-024-00129-w

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Alcaligenes faecalis is one of the most important and clinically significant environmental pathogens, increasing in importance due to its isolation from soil and nosocomial environments. The Gram-negative soil bacterium is associated with skin endocarditis, bacteremia, dysentery, meningitis, endophthalmitis, urinary tract infections, and pneumonia in patients. With emerging antibiotic resistance in A. faecalis, it has become crucial to understand the origin of such resistance genes within this clinically significant environmental and gut bacterium. In this research, we studied the impact of antibiotic overuse in poultry and its effect on developing resistance in A. faecalis. We sampled soil and faecal materials from five poultry farms, performed whole genome sequencing & analysis and identified four strains of A. faecalis. Furthermore, we characterized the genes in the genomic islands of A. faecalis isolates. We found four multidrug-resistant A. faecalis strains that showed resistance against vancomycin (MIC >1000 μg/ml), ceftazidime (50 μg/ml), colistin (50 μg/ml) and ciprofloxacin (50 μg/ml). From whole genome comparative analysis, we found more than 180 resistance genes compared to the reference sequence. Parts of our assembled contigs were found to be similar to different bacteria which included pbp1A and pbp2 imparting resistance to amoxicillin originally a part of Helicobacter and Bordetella pertussis. We also found the Mycobacterial insertion element IS6110 in the genomic islands of all four genomes. This prominent insertion element can be transferred and induce resistance to other bacterial genomes. The results thus are crucial in understanding the transfer of resistance genes in the environment and can help in developing regimes for antibiotic use in the food and poultry industry.

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First Case of Respiratory Infection in Rabbits Caused by Alcaligenes faecalis in Romania
Vlad Iorgoni, Ionica Iancu, Ionela Popa, Alexandru Gligor, Gabriel Orghici, Bogdan Sicoe, Corina Badea, Cristian Dreghiciu, Călin Pop, Timea Bochiș, Janos Degi, Luminita Costinar, Corina Pascu, Viorel Herman
Veterinary Sciences.2025; 12(1): 33. CrossRef

Review

Protective and pathogenic role of humoral responses in COVID-19: Uni Park , Nam-Hyuk Cho; J. Microbiol. 2022;60(3):268-275. Published online March 2, 2022; DOI: https://doi.org/10.1007/s12275-022-2037-8

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Since the advent of SARS-CoV-2 in Dec. 2019, the global endeavor to identify the pathogenic mechanism of COVID-19 has been ongoing. Although humoral immunity including neutralizing activity play an important role in protection from the viral pathogen, dysregulated antibody responses may be associated with the pathogenic progression of COVID-19, especially in high-risk individuals. In addition, SARS-CoV-2 spike-specific antibodies acquired by prior infection or vaccination act as immune pressure, driving continuous population turnover by selecting for antibody-escaping mutations. Here, we review accumulating knowledge on the potential role of humoral immune responses in COVID-19, primarily focusing on their beneficial and pathogenic properties. Understanding the multifaceted regulatory mechanisms of humoral responses during SARS-CoV-2 infection can help us to develop more effective therapeutics, as well as protective measures against the ongoing pandemic.

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Rise in broadly cross-reactive adaptive immunity against human β-coronaviruses in MERS-recovered patients during the COVID-19 pandemic
So-Hee Kim, Yuri Kim, Sangeun Jeon, Uni Park, Ju-Il Kang, Kyeongseok Jeon, Hye-Ran Kim, Songhyeok Oh, Ji-Young Rhee, Jae-Phil Choi, Wan Beom Park, Sang Won Park, Jeong-Sun Yang, Joo-Yeon Lee, Jihye Kang, Hyoung-Shik Shin, Yeonjae Kim, Seungtaek Kim, Yeon-
Science Advances.2024;[Epub] CrossRef
Distinctive Combinations of RBD Mutations Contribute to Antibody Evasion in the Case of the SARS-CoV-2 Beta Variant
Tae-Hun Kim, Sojung Bae, Sunggeun Goo, Jinjong Myoung
Journal of Microbiology and Biotechnology.2023; 33(12): 1587. CrossRef
Two years of COVID-19 pandemic: where are we now?
Jinjong Myoung
Journal of Microbiology.2022; 60(3): 235. CrossRef

Journal Articles

Mst1/2-ALK promotes NLRP3 inflammasome activation and cell apoptosis during Listeria monocytogenes infection: Aijiao Gao , Huixin Tang , Qian Zhang , Ruiqing Liu , Lin Wang , Yashan Liu , Zhi Qi , Yanna Shen; J. Microbiol. 2021;59(7):681-692. Published online April 20, 2021; DOI: https://doi.org/10.1007/s12275-021-0638-2

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Listeria monocytogenes (L. monocytogenes) is a Gram-positive intracellular foodborne pathogen that causes severe diseases, such as meningitis and sepsis. The NLR family pyrin domain-containing 3 (NLRP3) inflammasome has been reported to participate in host defense against pathogen infection. However, the exact molecular mechanisms underlying NLRP3 inflammasome activation remain to be fully elucidated. In the present study, the roles of mammalian Ste20- like kinases 1/2 (Mst1/2) and Anaplastic Lymphoma Kinase (ALK) in the activation of the NLRP3 inflammasome induced by L. monocytogenes infection were investigated. The expression levels of Mst1/2, phospho (p)-ALK, p-JNK, Nek7, and NLRP3 downstream molecules including activated caspase- 1 (p20) and mature interleukin (IL)-1β (p17), were upregulated in L. monocytogenes-infected macrophages. The ALK inhibitor significantly decreased the expression of p-JNK, Nek7, and NLRP3 downstream molecules in macrophages infected with L. monocytogenes. Furthermore, the Mst1/2 inhibitor markedly inhibited the L. monocytogenes-induced activation of ALK, subsequently downregulating the expression of p-JNK, Nek7, and NLRP3 downstream molecules. Therefore, our study demonstrated that Mst1/2-ALK mediated the activation of the NLRP3 inflammasome by promoting the interaction between Nek7 and NLRP3 via JNK during L. monocytogenes infection, which subsequently increased the maturation and release of proinflammatory cytokine to resist pathogen infection. Moreover, Listeriolysin O played a key role in the process. In addition, we also found that the L. monocytogenes-induced apoptosis of J774A.1 cells was reduced by the Mst1/2 or ALK inhibitor. The present study reported, for the first time, that the Mst1/2-ALK-JNK-NLRP3 signaling pathway plays a vital proinflammatory role during L. monocytogenes infection.

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PDE4B promotes JNK/NLRP3 activation in the nucleus pulposus and mediates intervertebral disc degeneration
Weixing Xu, Rana Dhar, Kaiyue Li, Danyang Zheng, Minxin He, Weiguo Ding, Long Xin, Bin Xu, Yuqing He, Qi Peng, Huifang Tang
Scientific Reports.2025;[Epub] CrossRef
IL-18 biology in severe asthma
Sarita Thawanaphong, Aswathi Nair, Emily Volfson, Parameswaran Nair, Manali Mukherjee
Frontiers in Medicine.2024;[Epub] CrossRef
TRAF6-TAK1-IKKβ pathway mediates TLR2 agonists activating “one-step” NLRP3 inflammasome in human monocytes
Mengdan Chen, Shi Yu, Yuhui Gao, Jiaxun Li, Xun Wang, Bin Wei, Guangxun Meng
Cytokine.2023; 169: 156302. CrossRef
ALK-JNK signaling promotes NLRP3 inflammasome activation and pyroptosis via NEK7 during Streptococcus pneumoniae infection
Xia Wang, Yan Zhao, Dan Wang, Chang Liu, Zhi Qi, Huixin Tang, Yashan Liu, Shiqi Zhang, Yali Cui, Yingying Li, Ruiqing Liu, Yanna Shen
Molecular Immunology.2023; 157: 78. CrossRef
Inflammasome activation by Gram-positive bacteria: Mechanisms of activation and regulation
A. Marijke Keestra-Gounder, Prescilla Emy Nagao
Frontiers in Immunology.2023;[Epub] CrossRef
Toxoplasma gondii profilin induces NLRP3 activation and IL-1β production/secretion in THP-1 cells
Hossein Pazoki, Hamed Mirjalali, Maryam Niyyati, Seyed Javad Seyed Tabaei, Nariman Mosaffa, Shabnam Shahrokh, Hamid Asadzadeh Ahdaei, Andreas Kupz, Mohammad Reza Zali
Microbial Pathogenesis.2023; 180: 106120. CrossRef
The Critical Role of Potassium Efflux and Nek7 in Pasteurella multocida-Induced NLRP3 Inflammasome Activation
Yu Wang, Zheng Zeng, Jinrong Ran, Lianci Peng, Xingping Wu, Chao Ye, Chunxia Dong, Yuanyi Peng, Rendong Fang
Frontiers in Microbiology.2022;[Epub] CrossRef
Coral and it's symbionts responses to the typical global marine pollutant BaP by 4D-Proteomics approach
Yuebin Pei, Shuai Chen, Yuting Zhang, Volovych Olga, Yuanchao Li, Xiaoping Diao, Hailong Zhou
Environmental Pollution.2022; 307: 119440. CrossRef
NEK7-Mediated Activation of NLRP3 Inflammasome Is Coordinated by Potassium Efflux/Syk/JNK Signaling During Staphylococcus aureus Infection
Ruiqing Liu, Yashan Liu, Chang Liu, Aijiao Gao, Lin Wang, Huixin Tang, Qiang Wu, Xia Wang, Derun Tian, Zhi Qi, Yanna Shen
Frontiers in Immunology.2021;[Epub] CrossRef

Whole genome analysis of Aspergillus sojae SMF 134 supports its merits as a starter for soybean fermentation: Kang Uk Kim , Kyung Min Kim , Yong-Ho Choi , Byung-Serk Hurh , Inhyung Lee; J. Microbiol. 2019;57(10):874-883. Published online June 27, 2019; DOI: https://doi.org/10.1007/s12275-019-9152-1

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Abstract PDF

Aspergillus sojae is a koji (starter) mold that has been applied for food fermentation in Asia. The whole genome of A. sojae SMF 134, which was isolated from meju (Korean soybean fermented brick), was analyzed at the genomic level to evaluate its potential as a starter for soybean fermentation. The genome size was 40.1 Mbp, which was expected to be composed of eight chromosomes with 13,748 ORFs. Strain SMF 134 had a total of 151 protease genes, among which two more leucine aminopeptidase (lap) genes were found in addition to the previously known lap1, and three γ-glutamyltranspeptidase (ggt) genes were newly identified. Such genomic characteristics of SMF 134 with many protease and flavor-related (lap and ggt) genes support its merits as a starter for soybean fermentation. In addition, this first complete genome of A. sojae will allow for further genetic studies to better understand the production of various enzymes, including proteases, LAPs, and GGTs, as well as other characteristics as a starter mold for soybean fermentation.

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Population Genomics of Aspergillus sojae is Shaped by the Food Environment
Kimberly L Acevedo, Elizabeth Eaton, Julia Leite, Shu Zhao, Katherine Chacon-Vargas, Colin M McCarthy, Dasol Choi, Samuel O’Donnell, Emile Gluck-Thaler, Jae-Hyuk Yu, John G Gibbons, Rebecca Zufall
Genome Biology and Evolution.2025;[Epub] CrossRef
Starter molds and multi-enzyme catalysis in koji fermentation of soy sauce brewing: A review
Yihao Liu, Guangru Sun, Jingyao Li, Peng Cheng, Qian Song, Wen Lv, Chunling Wang
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Lijie Zhang, Le Kang, Yan Xu, Yanbin Yin
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Characteristics of the soy sauce taste and koji enzyme profiles as affected by soybean traits
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Food Bioscience.2023; 53: 102776. CrossRef
Comparative proteome and volatile metabolome analysis of Aspergillus oryzae 3.042 and Aspergillus sojae 3.495 during koji fermentation
Jingyao Li, Bin Liu, Xiaojuan Feng, Mengli Zhang, Tingting Ding, Yue Zhao, Chunling Wang
Food Research International.2023; 165: 112527. CrossRef
CRISPR/Cas9 genome editing for comparative genetic analysis related to soy sauce brewing in Aspergillus sojae industrial strains
Takayuki Igarashi, Takuya Katayama, Jun-ichi Maruyama
Bioscience, Biotechnology, and Biochemistry.2023; 87(10): 1236. CrossRef
Untargeted metabolomic profiling of Aspergillus sojae 3.495 and Aspergillus oryzae 3.042 fermented soy sauce koji and effect on moromi fermentation flavor
Jingyao Li, Chengguo Sun, Zhanyu Shen, Yutong Tian, Fanghua Mo, Binghui Wang, Bin Liu, Chunling Wang
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Cesar A. Arreguin-Perez, Estefan Miranda-Miranda, Jorge Luis Folch-Mallol, Raquel Cossío-Bayúgar
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Arthur J. Lustig, Cecile Fairhead
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He-Jin Cho, Sung-Hun Son, Wanping Chen, Ye-Eun Son, Inhyung Lee, Jae-Hyuk Yu, Hee-Soo Park
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High molecular weight DNA extraction methods lead to high quality filamentous ascomycete fungal genome assemblies using Oxford Nanopore sequencing
Celine Petersen, Trine Sørensen, Klaus R. Westphal, Lavinia I. Fechete, Teis E. Sondergaard, Jens L. Sørensen, Kåre L. Nielsen
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Potential use of lactic acid bacteria Leuconostoc mesenteroides as a probiotic for the removal of Pb(II) toxicity: Young-Joo Yi , Jeong-Muk Lim , Suna Gu , Wan-Kyu Lee , Eunyoung Oh , Sang-Myeong Lee , Byung-Taek Oh; J. Microbiol. 2017;55(4):296-303. Published online March 31, 2017; DOI: https://doi.org/10.1007/s12275-017-6642-x

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It has been demonstrated that certain lactic acid bacteria (LAB) can sequester metal ions by binding them to their surfaces. In the present study, lead (Pb)-resistant LAB were isolated from kimchi, a Korean fermented food. A total of 96 different LAB strains were isolated, and 52 strains showed lead resistance. Among them, an LAB strain-96 (L-96) iden-tified as Leuconostoc mesenteroides showed remarkable Pb resistance and removal capacity. The maximum adsorption capacity of this strain calculated using the Langmuir isotherm was 60.6 mg Pb/g. In an in vivo experiment, young male mice were provided with water (A), Pb-water (B), or Pb-water+ L-96 (C) during puberty. Lower glutamate oxaloacetate trans-aminase (GOT) and glutamate pyruvate transaminase (GPT) levels in Pb-exposed male mice that received strain L-96 as a probiotic were suggestive of reduced hepatotoxicity. More-over, feces from mice treated with L-96 contained more Pb than feces from untreated mice. Increased Pb elimination likely reduced internal accumulation, and this hypothesis was supported by significantly lower Pb concentrations in kid-neys and testes of the mice treated with strain L-96. The mo-tility and ATP content of epididymal spermatozoa were par-tially restored if strain L-96 was administered. In conclusion, isolated L-96 LAB had lead-biosorption activity and effi-ciently detoxified lead-poisoned male mice, resulting in re-covering male reproductive function. These results suggest the potential use of LAB as a probiotic to protect humans from the adverse effects of Pb exposure.

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Aeration Effects on Metabolic Events during Sporulation of Bacillus thuringiensis: Mohammad H. Sarrafzadeh , Sabine Schorr-Galindo , Hyun-Joon La , Hee-Mock Oh; J. Microbiol. 2014;52(7):597-603. Published online June 28, 2014; DOI: https://doi.org/10.1007/s12275-014-3547-9

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The metabolism of Bacillus thuringiensis during its sporulation process was investigated under different concentrations of oxygen. At the beginning of sporulation, the aeration conditions were regulated to obtain different oxygen transfer rates (OTR) in four separate fermentations, representing interrupted, limited, non-limited, and saturated oxygenation, respectively. A higher OTR resulted in a higher pH, up to about 9 in the case of saturated oxygenation, while the interrupted oxygenation resulted in a significantly acidic culture. In contrast, the absence of oxygen resulted in rapid sporangia lysis and caused acidification of the medium, indicating a distinctly different sporangia composition and different metabolism. The bacterium also showed different CO2 production rates during sporulation, although amaximum point was observed in every case.With a higher OTR, the maximal value was observed after a longer time and at a lower value (40, 26, and 13 mmol/L/h for limited, non-limited, and saturated cases, respectively). Despite the exhaustion of glucose prior to the sporulation phase, the interrupted oxygenation resulted in acetate, lactate, and citrate in the medium with a maximum concentration of 4.8, 1.3, and 5.0 g/L, respectively. Notwithstanding, while the metabolic events differed visibly in the absence of oxygen, once sporulation was triggered, it was completed, even in the case of an interrupted oxygen supply.

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Solid-state fermentation of Bacillus thuringiensis var kurstaki HD-73 maintains higher biomass and spore yields as compared to submerged fermentation using the same media
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Production of an Endoinulinase from Aspergillus niger AUMC 9375, by Solid State Fermentation of Agricultural Wastes, with Purification and Characterization of the Free and Immobilized Enzyme: Manal M. Housseiny; J. Microbiol. 2014;52(5):389-398. Published online May 9, 2014; DOI: https://doi.org/10.1007/s12275-014-3561-y

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Two different substrates, sunflower (Helianthus annuus L.) tubers and lettuce (Lactuca sativa) roots, were tested. Using a mixture of both wastes resulted in higher production of endoinulinase than either waste alone. Also, ten fungal spe-cies grown on these substrates as inexpensive, carbon sour-ces were screened for the best production of endoinulinase activities. Of these, Aspergillus niger AUMC 9375 was the most productive, when grown on the mixture using a 6:1 w/w ratio of sun flower: lettuce, and yielded the highest levels of inulinase at 50% moisture, 30°C, pH 5.0, with seven days of incubation, and with yeast extract as the best nitrogen source. Inulinase was purified to homogeneity by ion-exchange chro-matography and gel-filtration giving a 51.11 fold purification. The mixture of sunflower tubers and lettuce roots has poten-tial to be an effective and economical substrate for inulinase production. Inulinase was successfully immobilized with an immobilization yield of 71.28%. After incubation for 2 h at 60°C, the free enzyme activity decreased markedly to 10%, whereas that of the immobilized form decreased only to 87%. A reusability test demonstrated the durability of the immo-bilized inulinase for 10 cycles and in addition, that it could be stored for 32 days at 4°C. These results indicate that this inulinase, in the immobilized form, is a potential candidate for large-scale production of high purity fructose syrups.

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Research Support, Non-U.S. Gov'ts

Solid State Production of Polygalacturonase and Xylanase by Trichoderma Species Using Cantaloupe and Watermelon Rinds: Saleh A. Mohamed , Abdulrahman L. Al-Malki , Jalaluddin A. Khan , Saleh A. Kabli , Saleh M. Al-Garni; J. Microbiol. 2013;51(5):605-611. Published online September 14, 2013; DOI: https://doi.org/10.1007/s12275-013-3016-x

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Different solid state fermentation (SSF) sources were tested such as cantaloupe and watermelon rinds, orange and banana peels, for the production of polygalacturonase (PG) and xylanase (Xyl) by Trichoderma harzianum and Trichoderma virens. The maximum production of both PG and Xyl were obtained by T. harzianum and T. virnes grown on cantaloupe and watermelon rinds, respectively. Time course, moisture content, temperature, pH, supplementation with carbon and nitrogen sources were optimized to achieve the maximum production of both PG and Xyl of T. harzianum and T. virens using cantaloupe and watermelon rinds, respectively. The maximum production of PG and Xyl of T. harzianum and T. virens was recorded at 4–5 days of incubation, 50–66% moisture, temperature 28–35°C and pH 6–7. The influence of supplementary carbon and nitrogen sources was studied. For T. harzianum, lactose enhanced PG activity from 87 to 120 units/g solid, where starch and maltose enhanced Xyl activity from 40 to 55–60 units/g solid for T. virnes. Among the nitrogen sources, ammonium sulphate, ammonium nitrate, yeast extract and urea increased PG activity from 90 to 110–113 units/g solid for T. harzianum. Similarly, ammonium chloride, ammonium sulphate and yeast extract increased Xyl activity from 45 to 55–70 units/g solid for T. virens.

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Enhancement of Butanol Tolerance and Butanol Yield in Clostridium acetobutylicum Mutant NT642 Obtained by Nitrogen Ion Beam Implantation: Xiao-Bo Liu , Qiu-Ya Gu , Xiao-Bin Yu , Wei Luo; J. Microbiol. 2012;50(6):1024-1028. Published online December 30, 2012; DOI: https://doi.org/10.1007/s12275-012-2289-9

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Abstract PDF

As a promising alternative biofuel, biobutanol can be produced through acetone/butanol/ethanol (ABE) fermentation. Currently, ABE fermentation is still a small-scale industry due to its low production and high input cost. Moreover, butanol toxicity to the Clostridium fermentation host limits the accumulation of butanol in the fermentation broth. The wild-type Clostridium acetobutylicum D64 can only produce about 13 g butanol/L and tolerates less than 2% (v/v) butanol. To improve the tolerance of C. acetobutylicum D64 for enhancing the production of butanol, nitrogen ion beam implantation was employed and finally five mutants with enhanced butanol tolerance were obtained. Among these, the most butanol tolerant mutant C. acetobutylicum NT642 can tolerate above 3% (v/v) butanol while the wide-type strain can only withstand 2% (v/v). In batch fermentation, the production of butanol and ABE yield of C. acetobutylicum NT642 was 15.4 g/L and 22.3 g/L, respectively, which were both higher than those of its parental strain and the other mutants using corn or cassava as substrate. Enhancing butanol tolerance is a great precondition for obtaining a hyperyield producer. Nitrogen ion beam implantation could be a promising biotechnology to improve butanol tolerance and production of the host strain C. acetobutylicum.

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NOTE] Ectopic Expression of Sweet Potato MuS1 Increases Acquired Stress Tolerance and Fermentation Yield in Saccharomyces cerevisiae: Il-Sup Kim , Sun-Young Shin , Sun-Hyung Kim , Ho-Sung Yoon; J. Microbiol. 2012;50(3):544-546. Published online June 30, 2012; DOI: https://doi.org/10.1007/s12275-012-2043-3

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The MuS1 gene is highly homologous to many stress-related proteins in plants. Here, we characterized whether a new candidate gene, MuS1, is related to multiple stress tolerance in yeast as it is in plants. Transgenic yeast strain expressing MuS1 were more resistant to hydrogen peroxide, menadione, high salinity, metals (i.e., cadmium, copper, iron, and zinc), ethanol, and lactic acid than wild-type strain transformed with a vector alone. In addition, the alcohol yield of the transgenic yeast strain was higher than that of the wild-type strain during the batch fermentation process. These results show that MuS1-expressing transgenic yeast strain exhibits enhanced alcohol yield as well as tolerance to abiotic stresses, especially metal stress.

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Journal Article

Purification and Characterization of a New L-Methioninase from Solid Cultures of Aspergillus flavipes: Ashraf S. A. El-Sayed; J. Microbiol. 2011;49(1):130-140. Published online March 3, 2011; DOI: https://doi.org/10.1007/s12275-011-0259-2

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L-Methioninase was purified to electrophoretic homogeneity from cultures of Aspergillus flavipes using anionexchange and gel filtration chromatography by 12.1 fold compared to the crude enzyme preparation. The purified enzyme had a molecular mass of 47 kDa under denaturing conditions and an isoelectric point of 5.8 with no structural glycosyl residues. The enzyme had optimum activity at pH 7.8 and pH stability from 6.8-8.0 at 35°C. The enzyme appeared to be catalytically stable below 40°C. The enzyme activity was strongly inhibited by DL-propargylglycine, hydroxylamine, PMSF, 2-mercaptoethanol, Hg2+, Cu2+, and Fe2+, with slight inhibition by Triton X-100. A. flavipes L-methioninase has a higher catalytic affinity towards L-methionine (Km, 6.5 mM and Kcat, 14.1 S-1) followed by a relative demethiolating activity to L-homocysteine (Km, 12 mM and Kcat, 9.3 S-1). The enzyme has two absorption maxima at 280 and 420 nm, typical of other PLP-enzymes. Apo-L-methioninase has the ability to reconstitute its structural catalytic state completely upon addition of 0.15 mM PLP. L-Methioninase has neither an appreciable effect on liver function, platelet aggregation, nor hemolysis of human blood. The purified L-methioninase from solid cultures of A. flavipes displayed unique biochemical and catalytic properties over the currently applied Pseudomonad enzyme.

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Research Support, Non-U.S. Gov'ts

Production, Partial Characterization, and Immobilization in Alginate Beads of an Alkaline Protease from a New Thermophilic Fungus Myceliophthora sp.: Letícia Maria Zanphorlin , Fernanda Dell Antonio Facchini , Filipe Vasconcelos , Rafaella Costa Bonugli-Santos , André Rodrigues , Lara Durães Sette , Eleni Gomes , Gustavo Orlando Bonilla-Rodriguez; J. Microbiol. 2010;48(3):331-336. Published online June 23, 2010; DOI: https://doi.org/10.1007/s12275-010-9269-8

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Abstract PDF

Thermophilic fungi produce thermostable enzymes which have a number of applications, mainly in biotechnological processes. In this work, we describe the characterization of a protease produced in solidstate (SSF) and submerged (SmF) fermentations by a newly isolated thermophilic fungus identified as a putative new species in the genus Myceliophthora. Enzyme-production rate was evaluated for both fermentation processes, and in SSF, using a medium composed of a mixture of wheat bran and casein, the proteolytic output was 4.5-fold larger than that obtained in SmF. Additionally, the peak of proteolytic activity was obtained after 3 days for SSF whereas for SmF it was after 4 days. The crude enzyme obtained by both SSF and SmF displayed similar optimum temperature at 50°C, but the optimum pH shifted from 7 (SmF) to 9 (SSF). The alkaline protease produced through solid-state fermentation (SSF), was immobilized on beads of calcium alginate, allowing comparative analyses of free and immobilized proteases to be carried out. It was observed that both optimum temperature and thermal stability of the immobilized enzyme were higher than for the free enzyme. Moreover, the immobilized enzyme showed considerable stability for up to 7 reuses.

Citations

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Kinetic Evaluation of Products Inhibition to Succinic Acid Producers Escherichia coli NZN111, AFP111, BL21, and Actinobacillus succinogenes 130ZT: Qiang Li , Dan Wang , Yong Wu , Maohua Yang , Wangliang Li , Jianmin Xing , Zhiguo Su; J. Microbiol. 2010;48(3):290-296. Published online June 23, 2010; DOI: https://doi.org/10.1007/s12275-010-9262-2

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Succinic acid is one of the platform compounds and its production via natural feedstocks has drawn worldwide concerns. To evaluate the inhibitory effects of fermentation products on the growth of Actinobacillus succinogenes 130ZT and Escherichia coli NZN111, AFP111, BL21, fermentations with addition of individual products in medium were carried out. The cell growth was inhibited when the concentrations of formate, acetate, lactate, and succinate were at range of 8.8-17.6 g/L, 10-40 g/L, 9-18 g/L, and 10-80 g/L, respectively. For these two species of bacteria, E. coli was more resistant to acid products than A. succinogenes, while both endured succinate rather than by-products. As a result of end product inhibition, succinate production yield by A. succinogenes decreased from 1.11 to 0.49 g/g glucose. Logistic and Monod mathematical models were presented to simulate the inhibition kinetics. The Logistic model was found more suitable for describing the overall synergistic inhibitory effects.

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Response of Saccharomyces cerevisiae to Stress-Free Acidification: Allen Kuan-Liang Chen , Cristy Gelling , Peter L. Rogers , Ian W. Dawes , Bettina Rosche; J. Microbiol. 2009;47(1):1-8. Published online February 20, 2009; DOI: https://doi.org/10.1007/s12275-008-0167-2

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Genome-wide transcriptional analysis of a Saccharomyces cerevisiae batch culture revealed that more than 829 genes were regulated in response to an environmental shift from pH 6 to pH 3 by added sulfuric acid. This shift in pH was not detrimental to the rate of growth compared to a control culture that was maintained at pH 6 and the transcriptional changes most strikingly implicated not up- but down-regulation of stress responses. In addition, the transcriptional changes upon acid addition indicated remodeling of the cell wall and central carbon metabolism. The overall trend of changes was similar for the pH-shift experiment and the pH 6 control. However, the changes in the pH 6 control were much weaker and occurred 2.5 h later than in the pH-shift experiment. Thus, the reaction to the steep pH decrease was an immediate response within the normal repertoire of adaptation shown in later stages of fermentation at pH 6. Artificially preventing the yeast from acidifying the medium may be considered physiologically stressful under the tested conditions.

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A Specific Short Dextrin-Hydrolyzing Extracellular Glucosidase from the Thermophilic Fungus Thermoascus aurantiacus 179-5: Ana Flavia Azevedo Carvalho , Aline Zorzetto Gonclves , Roberto da Silva , Eleni Gomes; J. Microbiol. 2006;44(3):276-283.; DOI: https://doi.org/2385 [pii]

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Abstract PDF: The thermophilic fungus Thermoascus aurantiacus 179-5 produced large quantities of a glucosidase which preferentially hydrolyzed maltose over starch. Enzyme production was high in submerged fermentation, with a maximal activity of 30 U/ml after 336 h of fermentation. In solid-state fermentation, the activity of the enzyme was 22 U/ml at 144 h in medium containing wheat bran and 5.8 U/ml at 48 h when cassava pulp was used as the culture medium. The enzyme was specific for maltose, very slowly hydrolyzed starch, dextrins (2-7G) and the synthetic substrate (α-PNPG), and did not hydrolyze sucrose. These properties suggest that the enzyme is a type II α-glucosidase. The optimum temperature of the enzyme was 70?. In addition, the enzyme was highly thermostable (100% stability for 10 h at 60? and a half-life of 15 min at 80?), and stable within a wide pH range.

Response of sheep rumen fermentation and microbial communities to feed infected with the endophyte Epichloë gansuensis as evaluated with rumen-simulating technology: Yaling Ma , Hucheng Wang , Chunjie Li; J. Microbiol. 2021;59(8):719-728.; DOI: https://doi.org/10.1007/s12275-021-1113-9

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Abstract PDF

Achnatherum inebrians, a perennial grass, is widely distributed in China. When infected by the endophyte Epichloë gansuensis, A. inebrians produces an abundance of alkaloids that enhance plant survival but are toxic to animals. Here we used in vitro fermentation to study the impact of endophyte- infected A. inebrians (E+) addition on rumen fermentation characteristics and on microbial community and diversity as assessed with amplicon sequencing technology. We examined E+ addition at five levels, E0, E25, E50, E75, and E100, corresponding to 0%, 25%, 50%, 75%, and 100% of the fermentation substrate, respectively. Both the fermentation characteristics and rumen microbial community structure differed significantly among treatments. E100 resulted in the highest values for pH, the Shannon index, Kiritimatiellaeota, and Lentisphaerae levels relative to the other treatments. In contrast, E25 was associated with higher levels of ammonia nitrogen, total volatile fatty acid, propionate, butyrate, isobutyrate, valerate, of the phyla Bacteroidetes and Firmicutes, and of the genus Prevotella_1, Succiniclasticum, Family_XIII_AD3011_group, Rikenellaceae_RC9_gut_group, Prevotellaceae_UCG-001, and Pyramidobacter as compared with other treatments. E50 resulted in the greatest values for the abundance-based coverage estimator (ACE) and the Chao1 index as compared with other treatments. E0 resulted in the greatest values for digestibility of dry matter, gas production, acetate, and Ruminobacter as compared with other treatments. This approach avoided animal toxicity experiments and confirmed that rumen fermentation characteristics and rumen microbiota were affected by E+ toxin. Therefore, E25 showed higher abundance in Prevotella_1, Prevotellaceae_ UCG-001, and Lachnospiraceae_XPB1014_group that implied they should play significant roles in E+ alkaloids degradation. And then, we can infer that rumen microorganisms should function as an antidote with respect to this poisoning reaction at moderate dietary percentages of E+.

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