Biochemical insights into the recombinant 10-deacetylbaccatin III-10-β-O-acetyltransferase enzyme from the Taxol-producing endophytic fungus Lasiodiplodia theobromae

2019 ◽  
Vol 366 (7) ◽  
Author(s):  
Balendra Sah ◽  
Kamalraj Subban ◽  
Chelliah Jayabaskaran
Keyword(s):  
Endophytic Fungi ◽  
Endophytic Fungus ◽  
Biosynthetic Pathway ◽  
Gel Filtration ◽  
Alternative Source ◽  
Lasiodiplodia Theobromae ◽  
Reading Frame ◽  
Baccatin Iii ◽  
Feasible Alternative ◽  
Near Future

ABSTRACT 10-deacetylbaccatin III-10-β-O-acetyltransferase (DBAT) is a key rate-limiting enzyme of the Taxol biosynthetic pathway, which is uncharacterized in Taxol-producing endophytic fungi. Here, an open reading frame of DBAT was cloned from the Taxol-producing endophytic fungus Lasiodiplodia theobromae (LtDBAT). The LtDBAT enzyme was heterologously expressed and purified by the affinity and gel filtration chromatography methods. The molecular weight of the purified protein was 49 kDa and its identity was confirmed by western blot. The purified LtDBAT enzyme was capable of catalyzing 10-deacetylbaccatin III into baccatin III, as shown by liquid chromatography–mass spectroscopy. The mass spectra of baccatin III were identical to the authentic baccatin III. The LtDBAT enzyme was characterized and the kinetic parameters of catalysis were determined. In addition, localization of LtDBAT was performed by using confocal microscopy and the result showed that the enzyme was localized in lipid droplets. Together, this study provides biochemical insights into the fungal recombinant DBAT enzyme that is involved in the Taxol biosynthetic pathway. In the near future, engineering of the LtDBAT enzyme and the Taxol biosynthetic pathway in endophytic fungi could be an eco-friendly and economically feasible alternative source for production of Taxol and its precursors.

scholarly journals Evaluation of spore inoculum and confirmation of pathway genetic blueprint of T13αH and DBAT from a Taxol-producing endophytic fungus

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Balabhadrapatruni V. S. K. Chakravarthi ◽  
Satpal Singh ◽  
Subban Kamalraj ◽  
Vijai Kumar Gupta ◽  
Chelliah Jayabaskaran
Keyword(s):  
Fusarium Solani ◽  
Endophytic Fungus ◽  
Competitive Inhibition ◽  
Liquid Media ◽  
Alternative Source ◽  
Natural Origin ◽  
Baccatin Iii ◽  
Enhanced Production ◽  
Significant Research ◽  
Selection For

AbstractTaxol (paclitaxel), a plant-derived anticancer drug, has been among the most successful anticancer drugs of natural origin. Endophytic fungi have been proposed as a prominent alternative source for Taxol and its intermediate Baccatin III, however the very low yields remain a hinderance to their commercial utilization. Significant research efforts towards this end are underway globally. Here, we report the results on our earlier reported Taxol-producing endophytic fungus, Fusarium solani from the standpoint of spores as seed inoculum and media selection for enhanced Taxol and baccatin III yields. Spores produced on M1D medium with 94.76% viability were used for further media optimization for Taxol and Baccatin III production in five different liquid media under static and shaker condition at different cultivation days. Taxol and Baccatin III when quantified through competitive inhibition enzyme immunoassay (CIEIA), showed maximum production at 136.3 µg L−1 and 128.3 µg L−1, respectively in the modified flask basal broth (MFBB) under shaking condition. Further, two important genes of this pathway, namely taxane 13α-hydroxylase (T13αH) and 10-deacetylbaccatin III-10-β-O-acetyltransferase (DBAT) have been identified in this fungus. These findings are hoped to assist in further manipulation and metabolic engineering of the parent F. solani strain towards the enhanced production of Taxol and baccatin III.

Antidiabetic Activity of Extract and Compounds from an Endophytic Fungus Nigrospora oryzae

Drug Research ◽  
2017 ◽  
Vol 67 (05) ◽  
pp. 308-311 ◽  
Author(s):  
Philip F. Uzor ◽  
Patience O. Osadebe ◽  
Ngozi J. Nwodo
Keyword(s):  
Abscisic Acid ◽  
Blood Sugar ◽  
Endophytic Fungi ◽  
Endophytic Fungus ◽  
Fasting Blood Sugar ◽  
Antidiabetic Activity ◽  
Diabetic Mice ◽  
Alternative Source ◽  
Fungal Extract ◽  
Nigrospora Oryzae

AbstractThe fungal extract as well as the 3 biosynthetic compounds, (S)-(+)-2-cis-4-trans-abscisic acid (1), , 7'-hydroxy-abscisic acid (2) and 4-des-hydroxyl altersolanol A (3) from the endophytic fungus, Nigrospora oryzae, isolated from Combretum dolichopetalum leaf were investigated for their antidiabetic potential.The antidiabetic activity was determined in alloxan-induced diabetic mice by monitoring their fasting blood sugar for 9 h.The extract and the compounds (1–3) significantly (p<0.001) reduced the fasting blood sugar of the diabetic mice.The present study has shown that the biosynthetic products of the endophytic fungus, N. oryzae, exhibited strong antidiabetic activity. It has further shown that endophytic fungi could be an alternative source of novel compounds for management of diabetes.

Bioinfomatics Analysis of the Anthocyanidin synthase Gene in Tree Peony

2014 ◽  
Vol 1010-1012 ◽  
pp. 1181-1184
Author(s):  
Yan Zhao Zhang ◽  
Yan Wei Cheng ◽  
Hui Yuan Ya ◽  
Chao Yun ◽  
Jian Ming Han ◽  
...  
Keyword(s):  
Plant Species ◽  
Structure Prediction ◽  
Biosynthetic Pathway ◽  
Resistance Breeding ◽  
Flower Color ◽  
Anthocyanin Synthesis ◽  
Tree Peony ◽  
Reading Frame ◽  
Transcriptome Database ◽  
Color Modification

Anthocyanin mainly responsible for flowers color in many plant species, it also accumulated in response to lots of environmental stress to reduce the damage to plant cell. Anthocyanin synthesis (ANS) protein is an important synthetase participated in anthocyanin biosynthetic pathway. In this study, we isolated the PsANS gene from transcriptome database built by our previous study. The PsANS gene contain an 1050bp open reading frame encoding 349 amino acid, phylogenetic analysis revealed that PsANS was segrated into a group with ANS from others plant species. Secondary and thri-dimension structure prediction also revealed that it may have similar function with ANS in others plant species. The identified PsANS gene would be helpful for further research in flower color modification and resistance breeding.

scholarly journals Aspergiloid I, an unprecedented spirolactone norditerpenoid from the plant-derived endophytic fungus Aspergillus sp. YXf3

2014 ◽  
Vol 10 ◽  
pp. 2677-2682 ◽  
Author(s):  
Zhi Kai Guo ◽  
Rong Wang ◽  
Wei Huang ◽  
Xiao Nian Li ◽  
Rong Jiang ◽  
...  
Keyword(s):  
Endophytic Fungus ◽  
Spectroscopic Data ◽  
Biosynthetic Pathway ◽  
Culture Broth ◽  
Ring System ◽  
X Ray Diffraction ◽  
X Ray ◽  
Anti Oxidant ◽  
Inhibitory Activities ◽  
Aspergillus Sp

An unusual C18 norditerpenoid, aspergiloid I (1), was isolated from the culture broth of Aspergillus sp. YXf3, an endophytic fungus derived from Ginkgo biloba. Its structure was unambiguously established by analysis of HRMS–ESI and spectroscopic data, and the absolute configuration was determined by low-temperature (100 K) single crystal X-ray diffraction with Cu Kα radiation. This compound is structurally characterized by a new carbon skeleton with an unprecedented 6/5/6 tricyclic ring system bearing an α,β-unsaturated spirolactone moiety in ring B, and represents a new subclass of norditerpenoid, the skeleton of which is named aspergilane. The hypothetical biosynthetic pathway for 1 was also proposed. The cytotoxic, antimicrobial, anti-oxidant and enzyme inhibitory activities of 1 were evaluated.

scholarly journals Recombinant Expression and Characterization of the Major β-Lactamase of Mycobacterium tuberculosis

1998 ◽  
Vol 42 (6) ◽  
pp. 1375-1381 ◽  
Author(s):  
Rama Kishan R. Voladri ◽  
David L. Lakey ◽  
Steven H. Hennigan ◽  
Barbara E. Menzies ◽  
Kathryn M. Edwards ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Recombinant Expression ◽  
Gel Filtration ◽  
Multidrug Resistant ◽  
Chromosomal Dna ◽  
Reading Frame ◽  
Major Mechanism ◽  
Dna Library ◽  
Class A ◽  
Resistant Tuberculosis

ABSTRACT New antibiotic regimens are needed for the treatment of multidrug-resistant tuberculosis. Mycobacterium tuberculosis has a thick peptidoglycan layer, and the penicillin-binding proteins involved in its biosynthesis are inhibited by clinically relevant concentrations of β-lactam antibiotics. β-Lactamase production appears to be the major mechanism by whichM. tuberculosis expresses β-lactam resistance. β-Lactamases from the broth supernatant of 3- to 4-week-old cultures of M. tuberculosis H37Ra were partially purified by sequential gel filtration chromatography and chromatofocusing. Three peaks of β-lactamase activity with pI values of 5.1, 4.9, and 4.5, respectively, and which accounted for 10, 78, and 12% of the total postchromatofocusing β-lactamase activity, respectively, were identified. The β-lactamases with pI values of 5.1 and 4.9 were kinetically indistinguishable and exhibited predominant penicillinase activity. In contrast, the β-lactamase with a pI value of 4.5 showed relatively greater cephalosporinase activity. An open reading frame in cosmid Y49 of the DNA library of M. tuberculosis H37Rv with homology to known class A β-lactamases was amplified from chromosomal DNA of M. tuberculosis H37Ra by PCR and was overexpressed in Escherichia coli. The recombinant enzyme was kinetically similar to the pI 5.1 and 4.9 enzymes purified directly from M. tuberculosis. It exhibited predominant penicillinase activity and was especially active against azlocillin. It was inhibited by clavulanic acid andm-aminophenylboronic acid but not by EDTA. We conclude that the major β-lactamase of M. tuberculosis is a class A β-lactamase with predominant penicillinase activity. A second, minor β-lactamase with relatively greater cephalosporinase activity is also present.

scholarly journals Isolation of Endophytic Fungi QPS 05 from Quercus phillyraeoides A. Gray and Its Potential for α-Glucosidase Inhibitory Activity

2018 ◽  
Vol 20 (1) ◽  
pp. 1-7
Author(s):  
Anastasia Wheni Indrianingsih ◽  
Amalia Indah Prihantini ◽  
Sanro Tachibana
Keyword(s):  
Fatty Acids ◽  
Endophytic Fungi ◽  
Inhibitory Activity ◽  
Endophytic Fungus ◽  
Inhibitory Concentration ◽  
Life Cycles ◽  
Ic50 Values ◽  
Quercus Phillyraeoides ◽  
Alternaria Sp ◽  
Harmful Effects

AbstractEndophytic fungi are the microorganisms that spend all or part of their life cycles within plant tissue without causing harmful effects on the plant. In this study, 14 endophytic fungus from Quercus phillyraeoides A. Gray were isolated. Alternaria sp. QPS 05, an endophytic fungi which was isolated from the stem of Q. phillyraeoides A. Gray showed the highest α-glucosidase inhibitory activity. Further separation of ethyl acetate extract from the fungus led to the isolation of active substance from hexane-soluble fraction which give fatty acids mixture consist of palmitic acid, oleic acid, linoleic acid and linolenic acid (1) strong inhibitory activity against α-glucosidase. Isolated fatty acids (1) had inhibitory concentration (IC50) values against Saccharomyces cerevisiae was 12.10 μg/mL. The results of the present study showed that endophytic fungus from Alternaria sp. QPS 05 potentially contained a rich source of natural antidiabetic medicine.

scholarly journals A Phenolic Compound of Endophytic Fungi Isolated from Stem of Syzygium aqueum and its Diuretic Activity

2021 ◽  
Vol 12 (5) ◽  
pp. 7040-7051
Keyword(s):  
Secondary Metabolites ◽  
Phenolic Compound ◽  
Endophytic Fungi ◽  
Endophytic Fungus ◽  
White Male ◽  
Urine Volume ◽  
Chloride Ions ◽  
Male Rats ◽  
Diuretic Activity ◽  
Fungus Extract

Guava (Syzygium aqueum), a typical Indonesian crop plant, has diuretic properties. Plants used as diuretic medicines can also be valuable sources for endophytic fungi containing diuretic drugs. The aims of this study were to isolate and identify endophytic fungi from S. aqueum stems, to isolate their secondary metabolites, and to conduct in vivo tests of the diuretic activity of an endophytic fungus extract on white male rats of the Wistar strain. Endophytic fungi were isolated by a direct plating method, and fungal isolates were identified molecularly. A phenolic compound was isolated by chromatography, and the chemical structure was identified spectroscopically. Doses of 100, 200, and 300 mg/kg BW were administered to white male rats divided into six groups (normal control, negative control, positive control, and the three treatment groups). Diuretic tests included urine volume; sodium, chloride, and calcium electrolyte levels; and pH. Phylogenetic analysis identified the endophytic fungal isolate as Trichoderma ghanense (isolation code SA1). The secondary metabolite isolated from the T. ghanense extract was a phenolic compound. Diuretic tests using the endophytic fungus extract showed the highest urine volume with a dose of 300 mg/kg BW. The diuretic test results showed an increase in urine volume and levels of sodium, potassium, and chloride ions in the urine. The extract of T. ghanense isolated from the stem of S. aqueum has the potential as initial therapy for hypertension because it contains phenolic secondary metabolites that show diuretic activity.

scholarly journals Bioactive Products from Endophytic Fungi of Sages (Salvia spp.)

Agriculture ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 543 ◽  
Author(s):  
Beata Zimowska ◽  
Monika Bielecka ◽  
Barbara Abramczyk ◽  
Rosario Nicoletti
Keyword(s):  
Medicinal Plants ◽  
Microbial Communities ◽  
Endophytic Fungi ◽  
New Technologies ◽  
Disruptive Innovation ◽  
Industrial Applications ◽  
Alternative Source ◽  
Biotechnological Potential ◽  
Sustainable Solutions ◽  
The World

In the aim of implementing new technologies, sustainable solutions and disruptive innovation to sustain biodiversity and reduce environmental pollution, there is a growing interest by researchers all over the world in bioprospecting endophytic microbial communities as an alternative source of bioactive compounds to be used for industrial applications. Medicinal plants represent a considerable source of endophytic fungi of outstanding importance, which highlights the opportunity of identifying and screening endophytes associated with this unique group of plants, widespread in diverse locations and biotopes, in view of assessing their biotechnological potential. As the first contribution of a series of papers dedicated to the Lamiaceae, this article reviews the occurrence and properties of endophytic fungi associated with sages (Salvia spp.).

scholarly journals 12-Membered Resorcylic Acid Lactones Isolated from Saccharicola bicolor, an Endophytic Fungi from Bergenia purpurascens

2015 ◽  
Vol 10 (12) ◽  
pp. 1934578X1501001 ◽  
Author(s):  
Da-Le Guo ◽  
Min Zhao ◽  
Shi-Ji Xiao ◽  
Bing Xia ◽  
Bo Wan ◽  
...  
Keyword(s):  
Endophytic Fungi ◽  
Endophytic Fungus ◽  
Spectroscopic Evidence ◽  
Resorcylic Acid Lactones

Two new resorcylic acid lactones, 13-hydroxyhidroresorcylide (1) and 12-hydroxyhidroresorcylide (2), along with four known congeners (3–6) were isolated from Saccharicola bicolor, an endophytic fungus from Bergenia purpurascens. Their structures were elucidated by interpretation of the spectroscopic evidence.

Biosynthesis of Porphyrinogens in Etiolated Euglena gracilis Z. I. Isolation and Purification of an Endogenous Factor Stimulating the Formation of Porphyrinogens

1988 ◽  
Vol 43 (5-6) ◽  
pp. 351-356 ◽  
Author(s):  
A. A. Juknat ◽  
D. Dörnemann ◽  
H. Senger
Keyword(s):  
Folic Acid ◽  
Euglena Gracilis ◽  
Biosynthetic Pathway ◽  
Gel Filtration ◽  
Supernatant Fraction ◽  
Isolation And Purification ◽  
Stable Factor ◽  
Heat Stable ◽  
Endogenous Compound ◽  
Euglena Gracilis Z

A low molecular weight, heat-stable factor has been purified from Euglena gracilis supernatant fraction by employing gel filtration, cation and anion exchange and paper chromatography. This endogenous compound stimulates porphobilinogenase (PBG-ase) (EC 4.3.1.8) activity, an enzyme of the porphyrin biosynthetic pathway. 10-7 ᴍ folic acid and 10-4 ᴍ 6-biopterin produced a significant activation, equivalent to 2-4 units of the purified factor. Elution patterns from the columns and fluorescence and UV absorption peaks suggest that this compound is a pteridine. This conclusion is further supported by the fact that both, folic acid and 6-biopterin can replace the action of the isolated factor on PBG-ase. The mechanism of stimulation is discussed.

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