Journal of Microbiology

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Tubulysin Production by the Dead Cells of Archangium gephyra KYC5002: Seohui Park, Chaehyeon Park, Yujin Ka, Kyungyun Cho; J. Microbiol. 2024;62(6):463-471. Published online June 13, 2024; DOI: https://doi.org/10.1007/s12275-024-00130-3

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Abstract: Archangium gephyra KYC5002 produces tubulysins during the death phase. In this study, we aimed to determine whether dead cells produce tubulysins. Cells were cultured for three days until the verge of the death phase, disrupted via ultrasonication, incubated for 2 h, and examined for tubulysin production. Non-disrupted cells produced 0.14 mg/L of tubulysin A and 0.11 mg/L of tubulysin B. Notably, tubulysin A production was increased by 4.4-fold to 0.62 mg/L and that of tubulysin B was increased by 6.7-fold to 0.74 mg/L in the disrupted cells. The same increase in tubulysin production was observed when the cells were killed by adding hydrogen peroxide. However, when the enzymes were inactivated via heat treatment of the cultures at 65 °C for 30 min, no significant increase in tubulysin production due to cell death was observed. Reverse transcription-quantitative polymerase chain reaction analysis of tubB mRNA revealed that the expression levels of tubulysin biosynthetic enzyme genes increased during the death phase compared to those during the vegetative growth phase. Our findings suggest that A. gephyra produces biosynthetic enzymes and subsequently uses them for tubulysin production in the cell death phase or during cell lysis by predators.

Saxibacter everestensis gen. nov., sp. nov., A Novel Member of the Family Brevibacteriaceae, Isolated from the North Slope of Mount Everest: Mao Tian, Shiyu Wu, Wei Zhang, Gaosen Zhang, Xue Yu, Yujie Wu, Puchao Jia, Binglin Zhang, Tuo Chen, Guangxiu Liu; J. Microbiol. 2024;62(4):277-284. Published online March 6, 2024; DOI: https://doi.org/10.1007/s12275-024-00108-1

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Abstract: We isolated and analyzed a novel, Gram-stain-positive, aerobic, rod-shaped, non-motile actinobacterium, designated as strain ZFBP1038(T), from rock sampled on the north slope of Mount Everest. The growth requirements of this strain were 10-37 °C, pH 4-10, and 0-6% (w/v) NaCl. The sole respiratory quinone was MK-9, and the major fatty acids were anteiso-C(15:0) and iso-C(17:0). Peptidoglycan containing meso-diaminopimelic acid, ribose, and glucose were the major cell wall sugars, while polar lipids included diphosphatidyl glycerol, phosphatidyl glycerol, an unidentified phospholipid, and an unidentified glycolipid. A phylogenetic analysis based on 16S rRNA gene sequences showed that strain ZFBP1038(T) has the highest similarity with Spelaeicoccus albus DSM 26341( T) (96.02%). ZFBP1038(T) formed a distinct monophyletic clade within the family Brevibacteriaceae and was distantly related to the genus Spelaeicoccus. The G + C content of strain ZFBP1038(T) was 63.65 mol% and the genome size was 4.05 Mb. Digital DNA-DNA hybridization, average nucleotide identity, and average amino acid identity values between the genomes of strain ZFBP1038(T) and representative reference strains were 19.3-25.2, 68.0-71.0, and 52.8-60.1%, respectively. Phylogenetic, phenotypic, and chemotaxonomic characteristics as well as comparative genome analyses suggested that strain ZFBP1038(T) represents a novel species of a new genus, for which the name Saxibacter gen. nov., sp. nov. was assigned with the type strain Saxibacter everestensis ZFBP1038(T) (= EE 014( T) = GDMCC 1.3024( T) = JCM 35335( T)).

Nano-encapsulation of naringinase produced by Trichoderma longibrachiatum ATCC18648 on thermally stable biopolymers for citrus juice debittering: Manal M. Housseiny , Heba I. Aboelmagd; J. Microbiol. 2019;57(6):521-531. Published online May 27, 2019; DOI: https://doi.org/10.1007/s12275-019-8528-6

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Abstract

Characteristics of naringinase nano-encapsulated forms on different carrier materials (chitosan and alginate polymers) were investigated in this study. Screening of twelve fungal isolates for naringinase production indicated that Trichoderma longibrachiatum was the most promising. Grapefruit rind was used as a substrate containing naringin for naringinase production. TEM micrographs showed that chitosan nano-capsules were applied for the production of morphologically homogeneous enzymatic nano-particles with high enzyme encapsulation efficiency, small asymmetric sizes (from 15.09 to 27.07 nm with the mean of 21.8 nm) and rough surfaces compared to nano-encapsulated naringinase in alginate which showed nano-particle size (from 33.37 to 51.01 nm with the mean of 43.03 nm). It was revealed that the highest naringinase activity was found in case of chitosan nano-capsule naringinase compared to alginate nano-capsule one. Thermogram analysis (TGA) showed that the free enzyme loses about 92% of its weight at approximately 110°C, while the nanoencapsulated ones show more stability at higher temperatures. Conclusively, the nano-capsulation process improves the kinetics and operational stability so could be useful as a debittering agent for various thermal processing applications in citrus juices industries which makes the fruit juice more acceptable and cost-effective to the consumer.

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Recent advancements in encapsulation of chitosan-based enzymes and their applications in food industry
Hongcai Zhang, Miaomiao Feng, Yapeng Fang, Yan Wu, Yuan Liu, Yanyun Zhao, Jianxiong Xu
Critical Reviews in Food Science and Nutrition.2023; 63(32): 11044. CrossRef
Alginate-based materials for enzyme encapsulation
Yilun Weng, Guangze Yang, Yang Li, Letao Xu, Xiaojing Chen, Hao Song, Chun-Xia Zhao
Advances in Colloid and Interface Science.2023; 318: 102957. CrossRef
Design and development of laboratory scale batch type device for debittering of bitter citrus juice
Arun Kumar Gupta, Muzamil Ahmad Rather, Poonam Mishra
Journal of Food Process Engineering.2023;[Epub] CrossRef
Current and emerging applications in detection and removal of bitter compounds in citrus fruit juice: A critical review
Arun Kumar Gupta, Subhamoy Dhua, Pratiksha, Vijay Kumar, Bindu Naik, Lembe Samukelo Magwaza, Khayelihle Ncama, Umezuruike Linus Opara, David Julian McClements, Poonam Mishra
Food Bioscience.2023; 55: 102995. CrossRef
Isolation and Molecular Characterization of the Naringinase Producing Micro-organisms for the Bio-transformation of Flavonoid
Ananda Sindhe, K. Lingappa
Journal of Pure and Applied Microbiology.2023; 17(1): 456. CrossRef
Preparation of Aspergillus niger 426 naringinases for debittering citrus juice utilization of agro-industrial residues
Fernanda de Oliveira, Tereza Cristina Luque Castellane, Marcelo Rodrigues de Melo, João Batista Buzato
International Microbiology.2022; 25(1): 123. CrossRef
Trends in the development of innovative nanobiocatalysts and their application in biocatalytic transformations
Elena Gkantzou, Alexandra V. Chatzikonstantinou, Renia Fotiadou, Archontoula Giannakopoulou, Michaela Patila, Haralambos Stamatis
Biotechnology Advances.2021; 51: 107738. CrossRef
Recent developments in enzyme immobilization technology for high-throughput processing in food industries
Asghar Taheri-Kafrani, Sara Kharazmi, Mahmoud Nasrollahzadeh, Asieh Soozanipour, Fatemeh Ejeian, Parisa Etedali, Hajar-Alsadat Mansouri-Tehrani, Amir Razmjou, Samaneh Mahmoudi-Gom Yek, Rajender S. Varma
Critical Reviews in Food Science and Nutrition.2021; 61(19): 3160. CrossRef
Polymers as Encapsulating Agents and Delivery Vehicles of Enzymes
Adejanildo da S. Pereira, Camila P. L. Souza, Lidiane Moraes, Gizele C. Fontes-Sant’Ana, Priscilla F. F. Amaral
Polymers.2021; 13(23): 4061. CrossRef

Trichoderma biodiversity in major ecological systems of China: Kai Dou , Jinxin Gao , Chulong Zhang , Hetong Yang , Xiliang Jiang , Jishun Li , Yaqian Li , Wei Wang , Hongquan Xian , Shigui Li , Yan Liu , Jindong Hu , Jie Chen; J. Microbiol. 2019;57(8):668-675. Published online May 23, 2019; DOI: https://doi.org/10.1007/s12275-019-8357-7

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Abstract

An investigation of Trichoderma biodiversity involving a large-scale environmental gradient was conducted to understand the Trichoderma distribution in China. A total of 3,999 isolates were isolated from forestry, grassland, wetland and agriculture ecosystems, and 50 species were identified based on morphological characteristics and sequence analysis of genetic markers. Trichoderma harzianum showed the largest proportion of isolates and the most extensive distribution. Hypocrea semiorbis, T. epimyces, T. konilangbra, T. piluliferum, T. pleurotum, T. pubescens, T. strictipilis, T. hunua, T. oblongisporum and an unidentified species, Trichoderma sp. MA 3642, were first reported in China. Most Trichoderma species were distributed in Jilin and Heilongjiang Provinces in northeast China and the fewest were distributed in Qinghai Province. Based on the division of ecological and geographic factors, forestry ecosystems and low-altitude regions have the greatest species biodiversity of Trichoderma.

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Genomic Characterization and Establishment of a Genetic Manipulation System for Trichoderma sp. (Harzianum Clade) LZ117
Jie Yang, Cristopher Reyes Loaiciga, Hou-Ru Yue, Ya-Jing Hou, Jun Li, Cheng-Xi Li, Jing Li, Yue Zou, Shuai Zhao, Feng-Li Zhang, Xin-Qing Zhao
Journal of Fungi.2024; 10(10): 697. CrossRef
Trichoderma Diversity in Mexico: A Systematic Review and Meta-Analysis
Hector Osvaldo Ahedo-Quero, Teodulfo Aquino-Bolaños, Yolanda Donají Ortiz-Hernández, Edgar García-Sánchez
Diversity.2024; 16(1): 68. CrossRef
Eleven new species of Trichoderma (Hypocreaceae, Hypocreales) from China
Rui Zhao, Ke-Yu Chen, Li-Juan Mao, Chu-Long Zhang
Mycology.2024; : 1. CrossRef
Screening of the Biocontrol Efficacy of Potent Trichoderma Strains against Fusarium oxysporum f.sp. ciceri and Scelrotium rolfsii Causing Wilt and Collar Rot in Chickpea
Ranjna Kumari, Vipul Kumar, Ananta Prasad Arukha, Muhammad Fazle Rabbee, Fuad Ameen, Bhupendra Koul
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Yunus Korkom, Ayhan Yıldız
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Nazia Manzar, Abhijeet Shankar Kashyap, Ravi Shankar Goutam, Mahendra Vikram Singh Rajawat, Pawan Kumar Sharma, Sushil Kumar Sharma, Harsh Vardhan Singh
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An in vitro study of the antifungal activity of Trichoderma virens 7b and a profile of its non-polar antifungal components released against Ganoderma boninense: Lee Pei Lee Angel , Mohd Termizi Yusof , Intan Safinar Ismail , Bonnie Tay Yen Ping , Intan Nur Ainni Mohamed Azni , Norman Hj Kamarudin , Shamala Sundram; J. Microbiol. 2016;54(11):732-744. Published online October 29, 2016; DOI: https://doi.org/10.1007/s12275-016-6304-4

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Abstract

Ganoderma boninense is the causal agent of a devastating disease affecting oil palm in Southeast Asian countries. Basal stem rot (BSR) disease slowly rots the base of palms, which radically reduces productive lifespan of this lucrative crop. Previous reports have indicated the successful use of Trichoderma as biological control agent (BCA) against G. boninense and isolate T. virens 7b was selected based on its initial screening. This study attempts to decipher the mechanisms responsible for the inhibition of G. boninense by identifying and characterizing the chemical compounds as well as the physical mechanisms by T. virens 7b. Hexane extract of the isolate gave 62.60% ± 6.41 inhibition against G. boninense and observation under scanning electron microscope (SEM) detected severe mycelial deformation of the pathogen at the region of inhibition. Similar mycelia deformation of G. boninense was observed with a fungicide treatment, Benlate® indicating comparable fungicidal effect by T. virens 7b. Fraction 4 and 5 of hexane active fractions through preparative thin layer chromatography (P-TLC) was identified giving the best inhibition of the pathogen. These fractions comprised of ketones, alcohols, aldehydes, lactones, sesquiterpenes, monoterpenes, sulphides, and free fatty acids profiled through gas chromatography mass spectrometry detector (GC/MSD). A novel antifungal compound discovery of phenylethyl alcohol (PEA) by T. virens 7b is reported through this study. T. virens 7b also proved to be an active siderophore producer through chrome azurol S (CAS) agar assay. The study demonstrated the possible mechanisms involved and responsible in the successful inhibition of G. boninense.

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Review of the state of the art of preparative thin-layer chromatography
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Research Support, Non-U.S. Gov'ts

Functional Analysis of a Subtilisin-like Serine Protease Gene from Biocontrol Fungus Trichoderma harzianum: Haijuan Fan , Zhihua Liu , Rongshu Zhang , Na Wang , Kai Dou , Gulijimila Mijiti , Guiping Diao , Zhiying Wang; J. Microbiol. 2014;52(2):129-138. Published online February 1, 2014; DOI: https://doi.org/10.1007/s12275-014-3308-9

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Abstract

The subtilisin-like serine protease gene ThSS45 has been cloned from Trichoderma harzianum ACCC30371. Its coding region is 1302 bp in length, encoding 433 amino acids, with a predicted protein molecular weight of 44.9 kDa and pI of 5.91. ThSS45 was shown by RT-qPCR analysis to be differentially transcribed in response to eight different treatments. The transcription of ThSS45 was up-regulated when grown in mineral medium, under carbon starvation, and nitrogen starvation, and in the presence of 1% root powder, 1% stem powder, and 1% leaf powder derived from Populus davidiana × P. bolleana (Shanxin poplar) aseptic seedlings. The highest increase in transcription approached 3.5-fold that of the control at 6 h under induction with 1% poplar root powder. The transcription of ThSS45 was also slightly up-regulated by 1% Alternaria alternata cell wall and 5% A. alternata fermentation liquid. Moreover, the analyses of coding and promoter regions of ThSS45 homologs indicated that serine protease may be involved in both mycoparasitism and antibiotic secretion. ThSS45 was cloned into the pGEX-4T-2 vector and then expressed in Escherichia coli BL21. The recombinant protein, with an expected molecular weight of approximately 69 kDa, was then purified. When transformant BL21-ss was induced with 1 mM IPTG for 6 h, the purified protease activity reached a peak of 18.25 U/ml at pH 7.0 and 40°C. In antifungal assays the purified protease obviously inhibited the growth of A. alternata mycelia.

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Subtilisin-like serine protease gene TghSS42 from Trichoderma ghanense ACCC 30153 was successfully expressed in Escherichia coli and recombinant protease rTghSS42 exhibited antifungal ability to five phytopathogens
HUIFANG ZHANG, NA WANG, YUCHENG WANG, JINJIE WANG, HONG ZHENG, ZHIHUA LIU
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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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Abstract

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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Cloning, Annotation and Expression Analysis of Mycoparasitism-Related Genes in Trichoderma harzianum 88: Lin Yao , Qian Yang , Jinzhu Song , Chong Tan , Changhong Guo , Li Wang , Lianhai Qu , Yun Wang; J. Microbiol. 2013;51(2):174-182. Published online April 27, 2013; DOI: https://doi.org/10.1007/s12275-013-2545-7

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Abstract: Trichoderma harzianum 88, a filamentous soil fungus, is an effective biocontrol agent against several plant pathogens. High-throughput sequencing was used here to study the mycoparasitism mechanisms of T. harzianum 88. Plate confrontation tests of T. harzianum 88 against plant pathogens were conducted, and a cDNA library was constructed from T. harzianum 88 mycelia in the presence of plant pathogen cell walls. Randomly selected transcripts from the cDNA library were compared with eukaryotic plant and fungal genomes. Of the 1,386 transcripts sequenced, the most abundant Gene Ontology (GO) classification group was “physiological process”. Differential expression of 19 genes was confirmed by real-time RT-PCR at different mycoparasitism stages against plant pathogens. Gene expression analysis revealed the transcription of various genes involved in mycoparasitism of T. harzianum 88. Our study provides helpful insights into the mechanisms of T. harzianum 88-plant pathogen interactions.

Construction of a Streptomyces lydicus A01 Transformant with a chit42 Gene from Trichoderma harzianum P1 and Evaluation of Its Biocontrol Activity against Botrytis cinerea: Qiong Wu , Linquan Bai , Weicheng Liu , Yingying Li , Caige Lu , Yaqian Li , Kehe Fu , Chuanjin Yu , Jie Chen; J. Microbiol. 2013;51(2):166-173. Published online April 27, 2013; DOI: https://doi.org/10.1007/s12275-013-2321-8

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Abstract: Streptomyces lydicus A01 and Trichoderma harzianum P1 are potential biocontrol agents of fungal diseases in plants. S. lydicus A01 produces natamycin to bind the ergosterol of the fungal cell membrane and inhibits the growth of Botrytis cinerea. T. harzianum P1, on the other hand, features high chitinase activity and decomposes the chitin in the cell wall of B. cinerea. To obtain the synergistic biocontrol effects of chitinase and natamycin on Botrytis cinerea, this study transformed the chit42 gene from T. harzianum P1 to S. lydicus A01. The conjugal transformant (CT) of S. lydicus A01 with the chit42 gene was detected using polymerase chain reaction (PCR). Associated chitinase activity and natamycin production were examined using the 3, 5-dinitrosalicylic acid (DNS) method and ultraviolet spectrophotometry, respectively. The S. lydicus A01-chit42 CT showed substantially higher chitinase activity and natamycin production than its wild type strain (WT). Consequently, the biocontrol effects of S. lydicus A01-chit42 CT on B. cinerea, including inhibition to spore germination and mycelial growth, were highly improved compared with those of the WT. Our research indicates that the biocontrol effect of Streptomyces can be highly improved by transforming the exogenous resistance gene, i.e. chit42 from Trichoderma, which not only enhances the production of antibiotics, but also provides a supplementary function by degrading the cell walls of the pathogens.

NOTE] The Microbial Population in the Air of Cultivation Facility of Oyster Mushrooms: Se Chul Chun , Yu Na Ahn , Sajid Mohamad Khan , Il Min Chung , Hyang Yoen Won , Chang Sung Jhune , Yool Jin Park; J. Microbiol. 2012;50(6):1053-1057. Published online December 30, 2012; DOI: https://doi.org/10.1007/s12275-012-2195-1

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Abstract: The microbial population in the air of mushroom cultivation facility was studied to understand the population structure and size depending on the cultivation methods and regions. The air contents of ten farmers’ oyster mushroom cultivation facilities in Kyunggi province were sampled. The results indicated that there was no difference in population size depending on the regions of mushroom cultivation. In addition, the population size of bacteria in the growth room was bigger than that of the cooling room and outside of the mushroom house, but the fungal population was similar in size between cultivation stages. With regard to population structure, Pseudomonas and Penicillium species were most frequently isolated from the air of oyster mushroom cultivation facility.

Journal Article

Characterization of Trichoderma reesei Endoglucanase II Expressed Heterologously in Pichia pastoris for Better Biofinishing and Biostoning: Sutanu Samanta , Asitava Basu , Umesh Chandra Halder , Soumitra Kumar Sen; J. Microbiol. 2012;50(3):518-525. Published online June 30, 2012; DOI: https://doi.org/10.1007/s12275-012-1207-5

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Abstract: The endoglucanase II of Trichoderma reesei is considered the most effective enzyme for biofinishing cotton fabrics and biostoning denim garments. However, the commercially available preparation of endoglucanase II is usually mixed with other cellulase components, especially endoglucanase I, resulting in hydrolysis and weight loss of garments during biofinishing and biostoning. We thus isolated the endoglucanase II gene from T. reesei to express this in Pichia pastoris, under the control of a methanol-inducible AOX1 promoter, to avoid the presence of other cellulase components. A highly expressible Mut+ transformant was selected and its expression in BMMH medium was found most suitable for the production of large amounts of the recombinant protein. Recombinant endoglucanase II was purified to electrophoretic homogeneity, and functionally characterized by activity staining. The specific activity of recombinant endoglucanase II was found to be 220.57 EU/mg of protein. Purified recombinant endoglucanase II was estimated to have a molecular mass of 52.8 kDa. The increase in molecular mass was likely due to hyperglycosylation. Hyperglycosylation of recombinant endoglucanase II secreted by P. pastoris did not change the temperature or pH optima as compared to the native protein, but did result in increased thermostability. Kinetic analysis showed that recombinant endoglucanase was most active against amorphous cellulose, such as carboxymethyl cellulose, for which it also had a high affinity.

Research Support, Non-U.S. Gov't

A Comparison of the Phenotypic and Genetic Stability of Recombinant Trichoderma spp. Generated by Protoplast- and Agrobacterium-Mediated Transformation: Rosa Elena Cardoza , Juan Antonio Vizcaino , Maria Rosa Hermosa , Enrique Monte , Santiago Gutierrez; J. Microbiol. 2006;44(4):383-395.; DOI: https://doi.org/2416 [pii]

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Abstract: Four different Trichoderma strains, T. harzianum CECT 2413, T. asperellum T53, T. atroviride T11 and T. longibrachiatum T52, which represent three of the four sections contained in this genus, were transformed by two different techniques: a protocol based on the isolation of protoplasts and a protocol based on Agrobacterium-mediated transformation. Both methods were set up using hygromycin B or phleomycin resistance as the selection markers. Using these techniques, we obtained phenotypically stable transformants of these four different strains. The highest transformation efficiencies were obtained with the T. longibrachiatum T52 strain: 65-70 transformants/μg DNA when transformed with the plasmid pAN7-1 (hygromycin B resistance) and 280 transformants/107 spores when the Agrobacterium-mediated transformation was performed with the plasmid pUR5750 (hygromycin B resistance). Overall, the genetic analysis of the transformants showed that some of the strains integrated and maintained the transforming DNA in their genome throughout the entire transformation and selection process. In other cases, the integrated DNA was lost.

Structure-antifungal activity relationships of cecropin a hybrid peptides against trichoderma sp.: Shin, Song Yub , Lee, Dong Gun , Lee, Sung gu , Kim, Kil Lyong , Lee, Myung Kyu , Hahm, Kyung Soo; J. Microbiol. 1997;35(1):21-24.

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Abstract: The hybrid peptides, CA-ME, CA-MA and CA-BO, with the N-terminal sequence 1-8 of cecropin A and the N-terminal sequences 1-12 of melittin, magainin 2 and bombinin, respectively, have more improved antibacterial activities. CA-MA was found to have stronger antifungal activity against Trichoderma sp than other hybrid peptides and their parental peptides. In order to elucidate the relationships between the peptide structure and antifungal activity, several analogues of CA-MA or CA-BO were also designed and synthesized by the solid phase method. Antifungal activity was measured against T. reesei and T. viride, and hemolytic activity was measured by a solution method against human red blood cells. The residue 16 of CA-MA, Ser, was found to be important for antifungal activity. When the residue was substituted with Leu, showed powerful antifungal activity was dramatically decreased. CA-MA, P1, P4 and P5 designed in this study showed powerful antifungal activity against T. reesei and T. viride with low hemolytic activity against human red blood cells. These hybrid peptides will be potentially useful model to further design peptides with powerful antifungal activity for the effective therepy of fungal infection and understand the mechanisms of antifungal actions of hybrid peptides.

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