Production, bioprocess optimization and anticancer activity of Camptothecin from Aspergillus terreus and Aspergillus flavus, endophytes of Ficus elastica

Elsayed, Abdelrahman H.; Khalaf, Salwa A.; Azez, Hany A.; Hussein, Hussein A.; EL-Moslamy, Shahira H.; Sitohy, Basel; El-Baz, Ashraf F.

doi:10.1016/j.procbio.2021.05.007

Cited by 27 publications

(54 citation statements)

References 66 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A dramatic decrease to the enzymatic activity by about 75% was measured at acidic conditions (pH 4.0), which is unlike the quite good enzymatic activity stability seen at slightly alkaline pHs (pH 10.0). Similar results were reported for TDS from T. baccata [68] and T. canadensis [69].…”

Section: Discussionsupporting

confidence: 88%

See 1 more Smart Citation

Purification and Biochemical Characterization of Taxadiene Synthase from Bacillus koreensis and Stenotrophomonas maltophilia

et al. 2021

Self Cite

View full text Add to dashboard Cite

Taxadiene synthase (TDS) is the rate-limiting enzyme of Taxol biosynthesis that cyclizes the geranylgeranyl pyrophosphate into taxadiene. Attenuating Taxol productivity by fungi is the main challenge impeding its industrial application; it is possible that silencing the expression of TDS is the most noticeable genomic feature associated with Taxol-biosynthetic abolishing in fungi. As such, the characterization of TDS with unique biochemical properties and autonomous expression that is independent of transcriptional factors from the host is the main challenge. Thus, the objective of this study was to kinetically characterize TDS from endophytic bacteria isolated from different plants harboring Taxol-producing endophytic fungi. Among the recovered 23 isolates, Bacillus koreensis and Stenotrophomonas maltophilia achieved the highest TDS activity. Upon using the Plackett–Burman design, the TDS productivity achieved by B. koreensis (18.1 µmol/mg/min) and S. maltophilia (14.6 µmol/mg/min) increased by ~2.2-fold over the control. The enzyme was purified by gel-filtration and ion-exchange chromatography with ~15 overall folds and with molecular subunit structure 65 and 80 kDa from B. koreensis and S. maltophilia, respectively. The chemical identity of taxadiene was authenticated from the GC-MS analyses, which provided the same mass fragmentation pattern of authentic taxadiene. The tds gene was screened by PCR with nested primers of the conservative active site domains, and the amplicons were sequenced, displaying a higher similarity with tds from T. baccata and T. brevifolia. The highest TDS activity by both bacterial isolates was recorded at 37–40 °C. The Apo-TDSs retained ~50% of its initial holoenzyme activities, ensuring their metalloproteinic identity. The activity of purified TDS was completely restored upon the addition of Mg2+, confirming the identity of Mg2+ as a cofactor. The TDS activity was dramatically reduced upon the addition of DTNB and MBTH, ensuring the implementation of cysteine-reactive thiols and ammonia groups on their active site domains. This is the first report exploring the autonomous robust expression TDS from B. koreensis and S. maltophilia with a higher affinity to cyclize GGPP into taxadiene, which could be a novel platform for taxadiene production as intermediary metabolites of Taxol biosynthesis.

show abstract

Section: Discussionsupporting

confidence: 88%

“…The purified TDS activity was dramatically reduced by upon the addition of DTNB and MBTH, respectively, ensuring the implementation of reactive thiols and ammonia groups on their active site domains [41][42][43]45,49,50,52,70,71]. Similar studies ensure the metalloproteinic identity of TDS from T. brevifolia and T. baccata [3,57,69].…”

Section: Discussionmentioning

confidence: 61%

Purification and Biochemical Characterization of Taxadiene Synthase from Bacillus koreensis and Stenotrophomonas maltophilia

et al. 2021

Self Cite

View full text Add to dashboard Cite

show abstract

“…The yield of Taxol by A. flavus has been optimized by the statistical experimental design of Plackett–Burman [ 18 , 28 ]. The minimum and maximum ranges of the selected parameters selected by Plackett–Burman design were summarized in Table 2 .…”

Section: Resultsmentioning

confidence: 99%

“…Placket-Burman design has been frequently used for optimization of the media component for fungal growth and production of bioactive secondary metabolites, evaluating the significant variables affecting Taxol production [ 18 , 20 , 46 ]. Choice of factor was based on media used in qualitative and quantitative screening.…”

Section: Methodsmentioning

confidence: 99%

Boosting the Anticancer Activity of Aspergillus flavus “endophyte of Jojoba” Taxol via Conjugation with Gold Nanoparticles Mediated by γ-Irradiation

Abdel-Fatah

El-Sherbiny

Khalaf

et al. 2022

Appl Biochem Biotechnol

View full text Add to dashboard Cite

Taxol production by fungi is one of the promising alternative approaches, regarding to the natural and semisynthetic sources; however, the lower yield and rapid loss of Taxol productivity by fungi are the major challenges that halt their further industrial implementation. Thus, searching for fungal isolates with affordable Taxol-production stability, in addition to enhance its anticancer activity via conjugation with gold nanoparticles, is the main objectives of this study. Twenty-four endophytic fungal isolates were recovered from the barks, twigs, and leaves of jojoba plant, among these fungi, Aspergillus flavus MW485934.1 was the most potent Taxol producer (88.6 µg/l). The chemical identity of the extracted Taxol of A. flavus was verified by the TLC, HPLC, HNMR, and FTIR analyses. The yield of Taxol produced by A. flavus was optimized by the response surface methodology (RSM) using Plackett–Burman (PBD) and faced central composite designs (FCCD). The yield of Taxol by A. flavus was increased by about 3.2 folds comparing to the control cultures (from 96.5 into 302.7 µg/l). The highest Taxol yield by was obtained growing A. flavus on a modified malt extract medium (g/l) (malt extract 20.0, peptone 2.0, sucrose 20.0, soytone 2.0, cysteine 0.5, glutamine 0.5, and beef extract 1.0 adjusted to pH 6.0) and incubated at 30 °C for 16 days. From the FCCD design, the significant variables affecting Taxol production by A. flavus were cysteine, pH, and incubation time. Upon A. flavus γ-irradiation at 1.0 kGy, the Taxol yield was increased by about 1.25 fold (375.9 µg/l). To boost its anticancer activity, the purified Taxol was conjugated with gold nanoparticles (AuNPs) mediated by γ-rays irradiation (0.5 kGy), and the physicochemical properties of Taxol-AuNPs composite were evaluated by UV–Vis, DLS, XRD, and TEM analyses. The IC50 values of the native-Taxol and Taxol-AuNPs conjugates towards HEPG-2 cells were 4.06 and 2.1 µg/ml, while the IC50 values against MCF-7 were 6.07 and 3.3 µg/ml, respectively. Thus, the anticancer activity of Taxol-AuNPs composite was increased by 2 folds comparing to the native Taxol towards HEPG-2 and MCF-7 cell lines. Also, the antimicrobial activity of Taxol against the multidrug resistant bacteria was dramatically increased upon conjugation with AuNPs comparing to authentic AuNPs and Taxol, ensuring the higher solubility, targetability, and efficiency of Taxol upon AuNPs conjugation.

show abstract

“…Herein, Oleic acid; is one of the unsaturated fatty acids, has been reported to exhibit various bioactivities such as anti-inflammatory, cancer preventive, hypochloestrolemic and dermatitigenic [ 101 ]. The compound 1,2-Benzenedicarboxylicacid, bis(2-ethylhexyl) ester was isolated from twigs of the dicot flowering plant Thevetia peruviana as a potential biomarker [ 102 ]. They added that this compound was proved to be a strong immunomodulatory B-cell stimulant.…”

Section: Resultsmentioning

confidence: 99%

Molecular and metabolic traits of some Egyptian species of Cassia L. and Senna Mill (Fabaceae-Caesalpinioideae)

et al. 2022

Self Cite

View full text Add to dashboard Cite

The genus Cassia and Senna have been classified under subfamily Caesalpinioideae of family Fabaceae (Leguminosae) of order Fabales. There is a scarce taxonomical studies of the genus Cassia and Senna inhabiting Egyptian environments, thus, the main objective of the current was to revise and authenticate the phylogenetic relationship between studied taxa of the species of the genera Cassia and Senna in Egypt using the recent tools of ITS barcoding, RAPD analysis and metabolic profiling, in comparing to the traditional taxonomical features. From the cluster analysis of the traditional 27 morphological characters, the studied taxa were categorized into two major clades with an average taxonomic distance of 4.3. The clade I include Cassia fistula, C. renigera, C. javanica L subsp. nodosa and C. roughiia that belongs to series Obolospermae, and C. grandis that belongs to series Grandes. The clade (II) includes Senna surattensis and S. alata at taxonomic level 3.6. The taxonomical description of the studied taxa was confirmed from the molecular analysis of ITS sequences and RAPD analysis. The ITS sequences of the tested plants species C. fistula L, C. grandis MD4, C. javanica subsp. nodosa MD7, C. roxburghii MD5, C. renigera MD5 were deposited at genbank with accession numbers MW367973, MZ960447, MW386305, MW326753 and MW32685, respectively. While, the ITS sequences of the S. surrattensis and S. alata were deposited into genbank accession # MD14 MW367670 and MD20 MW412635, respectively. Thus, from the molecular analysis, two clades were clearly separated into Clade I of Cassia and Clade II of Senna. The cluster I represented by C. fistula, C. renigera, C. roxburghii, and C. javanica sub nodosa, and the cluster II represented by S. alata and S. surattensis. From the PCA of RAPD, a clearly discrimination between the two Taxa was observed revealing the characteristic grouping of Cassia and Senna. The species Senna alata and Senna surattensis were grouped together, but the species of C. renigera, C. javanica, C. roxburghii and C. grandis was grouped on a distinct group. The separation of Cassia and Senna species into two clusters verify the segregation of the genus Cassia L. senso lato into two distinct genera namely Senna P. and Cassia L. The morphological, molecular traits of the studied plants were authenticated from the metabolic profiling by GC-MS analysis. Among the 23 identified metabolites, four compounds namely hexadecanoic acid, methyl ester, 9-Octadecenoic acid (Z)-ethyl ester and Vitamin E were detected with fluctuated concentrations, among C. fistula, C. grandis, C. javanica subsp. nodosa and C. roxburghii. Conclusively, the traditional morphological features, molecular barcoding using ITS sequences, RAPD analysis and metabolic traits by GC-MS analysis, authenticates the taxonomical diversity of the genus Cassia and Senna.

show abstract

Production, bioprocess optimization and anticancer activity of Camptothecin from Aspergillus terreus and Aspergillus flavus, endophytes of Ficus elastica

Cited by 27 publications

References 66 publications

Purification and Biochemical Characterization of Taxadiene Synthase from Bacillus koreensis and Stenotrophomonas maltophilia

Purification and Biochemical Characterization of Taxadiene Synthase from Bacillus koreensis and Stenotrophomonas maltophilia

Boosting the Anticancer Activity of Aspergillus flavus “endophyte of Jojoba” Taxol via Conjugation with Gold Nanoparticles Mediated by γ-Irradiation

Molecular and metabolic traits of some Egyptian species of Cassia L. and Senna Mill (Fabaceae-Caesalpinioideae)

Contact Info

Product

Resources

About