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Cephalotaxus mannii Hook. f. is a rare evergreen conifer native listed in the International Union for Conservation of Nature (IUCN) Red List, which is utilized for leukemia treatment. Although endophytic fungi from C. mannii was reported before, their cytotoxic property has not been revealed yet. In the present study, a total of 7 endophytic fungi were isolated from C. mannii collected in Ha Giang province, Vietnam, among which the isolate WQF18 was active against 5 tested pathogens with inhibition zones ranging from 18.0 ± 0.7 to 25.0 ± 0.4 mm. In addition, only ethyl acetate extract of isolate WQF18 showed cytotoxicity on A549 and MCF7 cell lines with IC50 values of 69.6 ± 2.3 µg/mL and 78.6 ± 1.6 µg/mL, respectively. PCR-based molecular screening revealed that the positive hits for both 10-deacetylbaccatin III-10-O-acetyl transferase (dbat) and taxadiene synthase (ts) genes involved in paclitaxel biosynthesis were only observed in the WQF18 isolate. Based on morphological and molecular identification, the WQF18 isolate was identified as Talaromyces wortmannii WQF18. The presence of paclitaxel in T. wortmannii WQF18 was further confirmed by dbat sequence alignment, phenotypic, and HPLC-DAD analysis. To the best of our knowledge, this is the first report demonstrating the paclitaxel-producing capability of endophytic fungi from C. mannii. These findings provide a new platform for deciphering paclitaxel biosynthesis of endophytic fungi from non-Taxus plants and further paclitaxel production.

Cephalotaxus mannii Hook.f is a rare medicinal plant used for the treatment of leukemia, however, its fungal endophytes with antibacterial and anticancer properties have not been exploited yet. In this study, six fungal isolates were recovered from roots of C. mannii collected in Ha Giang province, Vietnam. Among them, ethyl acetate crude extract of strain WQF5 exhibited strong antibacterial activity against 5 tested pathogens with inhibition zones ranging from 13.0 ± 0.5 mm to 20.0 ± 0.4 mm. Sulforhodamine B assay showed that WQF5 extract possessed the most potent cytotoxic activity against lung cancer A549 and breast cancer MCF7 cell lines with IC50 values of 6.9 ± 0.7 µg/mL and 23.1 ± 1.9 µg/mL, respectively. PCR-based molecular marker screening revealed that the positive hits for essential genes encoding 10-deacetylbaccatin III-10-O-acetyltransferase (dbat) and C-13 phenylpropanoyl side chain-CoA acyltransferase (bapt) involved in paclitaxel production were found in the fungal isolate WQF5. In addition, isolate WQF5, identified as Fusarium fujikuroi by morphological and ITS analysis, also produced paclitaxel as shown by HPLC-DAD analysis. This is the first report of bioprospecting endophytic fungi isolated from C. mannii, in which the capability of producing paclitaxel of endophytic fungi was also proved for the first time. These findings addressed a potent candidate for paclitaxel production and provided excellent material for further investigations of how endophytic fungi from non-Taxus plant species synthesize paclitaxel.

Lactic acid bacteria have been advocated as probiotics to replace antibiotic growth promoters, improve growth performance, and reduce cost production in swine farming. The aim of this study is to identify and evaluate the probiotic traits of strain LCN13 isolated from traditionally fermented feed for swine. Thirty-five bacterial isolates with different morphological characteristics were isolated, among which 9 isolates showed the ability to produce lactic acid and antibacterial activity against pathogenic bacteria. Among them, isolate LCN13 exhibited a strong capacity to produce lactic acid (18.5 ± 0.31 g/L), inhibited gastrointestinal pathogens such as Salmonella typhimurium ATCC 14028 (18.3 ± 1.52 mm), Escherichia coli ATCC 11105 (24.7 ± 2.14 mm), and Staphylococcus epidermidis ATCC 35984 (31.6 ± 2.93 mm), and produced 182 ng/mL folate and 233 ng/mL riboflavin as measured by LC-MS/MS after 24 hours of incubation. Based on morphological, biochemical, and 16S rRNA gene analysis, the isolate LCN13 was identified as Lactobacillus plantarum. Phenotypic analysis revealed that L. plantarum LCN13 showed remarkable resistance to 1.2% ox-bile salt, 2.0 mM H2O2, and pH 3.0. In addition, the ability to produce high levels of folate (253.6 ± 10.7 ng/mL) and riboflavin (312.0 ± 12.2 ng/mL) after 48 hours was exploited for the first time in the L. plantarum. Taken together, L. plantarum LCN13 might serve as a potential probiotic candidate for animal farming.

B-vitamins are micronutrients that play an important role in various cellular processes of organisms, which are only synthesized by plants, yeasts, and bacteria. Since animals and humans lack the ability to synthesize B-vitamins, supplements of vitamins from dietary and the B-vitamin producing bacteria are required. In this study, we, for the first time, shed some light on biosynthetic pathways involved in folate (vitamin B9), riboflavin (vitamin B2), and biotin (vitamin B7) production in Bacillus velezensis VTX20. The genome-wide comparison revealed that B. velezensis VTX20 shared high similarities with B. tequilensis KCTC 13622, B. subtilis 168, B. amyloliquefaciens DSM 7. Genomic analysis revealed the presence of a complete folate biosynthesis pathway in which some core components were not found in most Bacillus species. Moreover, strain VTX20 also had the metabolic pathways for riboflavin and biotin that are important probiotic traits. These results highlighted that B. velezensis VTX20 is a producer of B-vitamins, which can be applied further in the agricultural biotechnology industry.

Four new

Vietnam is recognized as one of the countries with the high diversity of medicinal plant species in the world, nevertheless little is known about the distribution, diversity and biological activity of endophytic actinomycetes associated with host plants. The present study aimed to evaluate antimicrobial and cytotoxic activities of four endophytic Streptomyces strains including Streptomyces sp. HBQ75, HBQ87, HBQ102 and HBQ104 isolated from different organs (roots, stems or leaves) of Cinnamomum cassia Presl. Analysis of 16S rRNA gene sequences and the phylogenetic tree assigned them to four different Streptomyces species as follow Streptomyces fulvissimus HBQ75, Streptomyces parvulus HBQ87, Streptomyces pratensis HBQ102 and Streptomyces ribosidificus HBQ104. These strains exhibited broad antimicrobial spectrum against at least five out of nine pathogens tested, among them S. parvulus HBQ87 showed the best activity (inhibition zones >20 mm). Interestingly, S. parvulus HBQ87 carried all three genes (pks-I, pks-II and nrps) encoding for polyketide synthase or non-ribosomal peptide synthetase enzymes involved in biosynthesis of secondary metabolites, while the remaining strains only possessed one or two genes. All the Streptomyces strains were positive for the anthracyclines-like antibiotic activity. The cell-free supernatants of S. parvulus HBQ87 revealed remarkable inhibitory effects against all three human cacinomar cell lines including hepatoma Hep3B, breast adenocarcinoma MCF7 and lung cancer A549 cells at both concentrations tested (30 µg/mL and 100 µg/mL), while S. fulvissimus HBQ75 and S. pratensis HBQ102 were active against only Hep3B and MCF7 cells. In conclusion, the phenotypic and genotypic features of the four endophytic Streptomyces strains suggest that they have a capacity to produce different broad-spectrum secondary metabolites. Among them, S. parvulus HBQ87 could be the most potential candidate for the production of important antimicrobial and antitumor compounds.

The Aspergillus fungi have been an important source of natural products that are useful for exploration in medicine, agriculture and industry. In our continuous investigation to search for new antimicrobial agents from marine-derived fungi, one new phomaligol A2 (1), together with three known compounds, wasabidienone E (2), aspertetranone D (3) and mactanamide (4), were obtained from the EtOAc extract of the culture medium of the marine-derived fungus Aspergillus flocculosus (A. flocculosus) 01NT.1.1.5 isolated from the sponge Stylissa sp. at Nhatrang Bay, Vietnam. Their chemical structures were elucidated by analysis of 1D and 2D NMR and mass spectroscopic data, as well as by comparison of the corresponding data to those previously reported in the literature. Furthermore, the aim of this study was also to evaluate the antimicrobial activity of these compounds against pathogenic microbes including Escherichia coli (E. coli) ATCC 25922, Pseudomonas aeruginosa (P. aeruginosa) ATCC 27853, Staphylococcus aureus (S. aureus) ATCC 25923, Bacillus cereus (B. cereus) ATCC 11778, Streptococcus faecalis (S. faecalis) ATCC 19433, Listeria monocytogenes (L. monocytogenes) ATCC 19111, and Candida albicans (C. albicans) ATCC 10231. Among the compounds, 1-3 were inhibitory on the growth of the yeast C. albicans with minimum inhibitory concentration (MIC) value of 16 μg/mL, which was more potent than amoxicillin and cefotaxime (MIC > 256 μg/mL), antimicrobial drugs as positive references. Moreover, compounds 1-4 were also found to be active against other pathogens including P. aeruginosa and S. faecalis with MIC values of 16 μg/mL and 32 μg/mL, respectively. Compound 4 had no inhibitory activity against L. monocytogenes, whereas compounds 1-3 had ability to against this strain with MICs of 32 to 64 μg/mL. Four of tested compounds exhibited antibacterial activity against B. cereus and E. coli with MIC values of 64-128 μg/mL. This is the first report about these compounds with antimicrobial activity obtained from marine fungus A. flocculosus isolated in Vietnam.

Endophytic bacteria reside in the intercellular spaces in different plant tissues without causing damage. This study aimed to select endophytic bacterial strains from Man Trau grass (Eleusine indica) that are able to inbibit the fungus A. alternata causing stem rot disease on pitaya. The result showed that one fungal isolate was pathogenic and caused stem rot disease on dragonfruit. The analysis results based on morphological characteristics and ITS region sequence indicated that the isolated fungal strain is 100% identical to A. alternata. In addition, a total of 16 endophytic bacterial strains were isolated from Man Trau grass samples in Binh Thuan and screened for antifungal activity. Ten bacterial strains with antagonistic activity against A. alternata causing stem rot disease on pitaya were detected. Among those, strain MT47 showed the strongest antifungal activity with an efficiency of 36.67%. Moreover, the result also presented that the antagonistic strain MT47 pocesses the ability of biofilm formation significantly higher than the control. The results suggest that these endophytic bacteria can be used in biocontrol of the stem rot disease on pitaya.

Currently, antibiotic resistance in pathogenic bacteria is a significant clinical problem with the increase of deseases and a serious public health concern. Thus, the identification of new antimicrobial agents, especially the secondary metabolites products by endophytic actinobacteria from medical plants could be promising sources of biologically active compounds in medical fields. This study focused on identification and evaluation of antimicrobial activity against pathogens; genes involved in their secondary metabolisms, and screening of anthracycline producing capacity (mainly presented in anti-cancer antibiotics) of YBQ75 isolated from Cinnamomum cassia Presl. plants in Yen Bai province. Based on manual of bacterial classification, method in International Streptomyces Project (ISP) and the 16S rRNA gene sequence (GenBank Acc. No. KR814822), the endophytic actinomycetes YBQ75 was named Streptomyces cavourensis YBQ75 with 100% identity. The strain S. cavourensis YBQ75 showed the remarkable antibacterial activities against 5 tested pathogens (Salmonella enterica ATCC 14028 (22.0 mm); Pseudomonas aeruginosa CNLM (19.3 mm); Staphylococcus epidermidis ATCC 12228 (19.3 mm); Enterobacter aerogenes ATCC 13048 (17.7 mm) and Proteus vulgaris CNLM (16.3 mm)) in the total of 9 tested pathogens. The detection of genes involved in antibiotic synthesis indicated that the strain S. cavourensis YBQ75 consists of all three genes related to antibiotic synthesis including polyketide synthase (pks-I) type I, polyketide synthase type II (pks-II) and nonribosomal peptide synthetase (nrps). Premarilly result showed that the strain S. cavourensis YBQ75 also present as an anthracycline productive actinomycetes. The resutls demonstrated that the endophytic actinomycetes S. cavourensis YBQ75 from medical plants could be promising sources for the production of antibiotics and anthracycline anticancer compounds.

The marine environment is an extremely complex ecosystem and contains a broad spectrum of fungal diversity. Marine fungi have been shown to be tremendous sources for new and biologically active secondary metabolites. The present study aims to isolate and screen antimicrobial properties of 100 fungus strains from different marine sources including seaweeds, soft corals, sponges and sediment collected at Nha Trang Bay, Vietnam. In preliminary experiments, the crude extracts of these fungal isolates with ethyl acetate were screened for their antimicrobial activity against the human microbial pathogens including Bacillus cereus ATCC 11778, Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853, Listeria monocytogenes ATCC 19111, Streptococcus faecalis ATCC 19433 and Candida albicans ATCC 10231 by the disc diffusion method. Among the 100 isolates, 59 strains exhibited antimicrobial activity against at least two tested pathogens, that 57% against S. aureus, 50% against L. monocytogenes, 49% against B. cereus, 45% against S. faecalis, 7% against E. coli, 5% against C. albicans, and only 2% against P. aeruginosa. The present study has revealed the presence of high numbers of marine fungi from Nha Trang waters having antimicrobial properties and they need to be investigated further for natural bioactive products.

The biosynthesis of compounds with antibiotic activity produced by marine fungi, strongly depends on their growth conditions. A good understanding of the role of culture conditions in the biosynthesis of metabolites may lead to better exploitation of microbial metabolites. In this study, the influence of culture conditions including incubation period, initial pH and salinity on antimicrobial activity and secondary metabolites production of marine fungus 01NT.1.1.5 was investigated. This isolate, obtained from sponge Stylissa sp. in Nha Trang Bay, exhibited a broad spectrum of in vitro antimicrobial activity to Bacillus cereus ATCC 11778, Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, Listeria monocytogenes ATCC 19111, Streptococcus faecalis ATCC 19433 and Candida albicans ATCC 10231. According to morphological characteristics and sequence analysis of 28S rDNA, the fungus was identified as Aspergillus flocculosus. The results indicated that antimicrobial activity and metabolite amount were highest when the fungus was cultivated in rice medium with incubation period of 20 days. The optimum salinity of 35 g/L and initial pH of 6.0 were found for the maximum antibiotic production. The colony growth, antimicrobial activity and production of secondary metabolites of the strain A. flocculosus 01NT.1.1.5 varied depending on salt concentrations and initial pH of medium. Particularly, extract of this fungus only showed activity against C. albicans when it was cultured in medium with 30-35 g/L salinity and initial pH 4.0-8.0. The results indicate that salinity and initial pH along with cultivation period are important factors influencing antimicrobial activity and secondary metabolites of A. flocculosus 01NT.1.1.5, and might be for other marine fungi.

Biotransformation of secondary metabolites from medicinal plants by bacteria has been studied extensively in the recent years. One of the potential sources for screening bacteria is plant endophytic bacteria. In this study, we isolated and screened endophytic bacteria for transformation of ginsenoside Rb1 of Ngoc Linh ginseng collected at village number 2, Tra Linh commune, Nam Tra My district, Quang Nam province. There are 45 strains isolated form rhizobium (24 strains), petioles (8 strains) and leaves (13 strains). After screening, there are 27 strains positive with β-glucosidase test, an enzyme which catalyses the hydrolysis of terminal non-reducing residues in β-glucosides-aglycone linkage of ginsenoside Rb. By evaluating the β-glucosidase activity and identification via 16S rRNA sequence, we choosed four high β-glucosidase acitivity strains SVK32 (Enterobacter sp.); SVK34 (Serratia sp.); SVK37 (Ochrobactrum sp.) and SVK44 (Arthrobacter sp.) for futher study on biotransformating of ginsenoside Rb1 into ginsenoside Rd and Rg3.