Decolorisation and detoxification of Direct Blue-15 by a bacterial consortium

Studies were carried out on decolorisation and biotransformation of the dye Direct Blue-15 into 3,3'-dimethoxybenzidine (O'-dianisidine) and a sulphonated derivative by a five-member bacterial consortium. Chromatographic studies revealed further complete biodegradation of 3,3'-dimethoxybenzidine coupled with release of ammonia, but the recalcitrant sulphonated derivative persisted. The microorganisms identified in the mixed consortium by 16S rDNA sequence analysis were Alcaligenes faecalis, Sphingomonas sp. EBD, Bacillus subtilis, Bacillus thuringiensis and Enterobacter cancerogenus. The cytotoxicity data showed a significant reduction in the toxicity (P<0.001) of the degraded dye as evidenced from the number of viable human polymorphonuclear leukocyte cells present.     

Modulatory effect of Gynandropsis gynandra L. on glucose metabolizing enzymes in aflatoxin B1-induced hepatocellular carcinoma in rats

The modulation of glucose-metabolizing enzymes activities play a vital role in the depletion of energy metabolism and leads to inhibition of cancer growth. In the present study, the effect of Gynandropsis gynandra L. extract on aflatoxin B1 (AFB1)-induced hepatocellular carcinoma (HCC) was studied on glucose-metabolizing enzymes in rats. A significant increase (p < 0.001) in the activities of the key glycolytic enzymes viz., hexokinase and phosphoglucoisomerase, with a significant decrease (p < 0.001) in the gluconeogenic enzymes glucose-6-phosphatase and fructose-1,6-bisphosphatase were observed in HCC-bearing rats, when compared with the control. Administration of G. gynandra extract caused a significant decrease in the activities of glycolytic enzymes and an increase in the gluconeogenic enzymes activities to near normal values. Thus, findings suggest the G. gynandra extract has a definite modulating role on the key enzymes of glucose metabolism in HCC. The modulatory effect may be due to the phytoactive constituents present in the extract of G. gynandra.     

The occurrence and control of nitric oxide generation by the plant mitochondrial electron transport chain

The plant mitochondrial electron transport chain (ETC) is bifurcated such that electrons from ubiquinol are passed to oxygen via the usual cytochrome path or through alternative oxidase (AOX). We previously showed that knockdown of AOX in transgenic tobacco increased leaf concentrations of nitric oxide (NO), implying that an activity capable of generating NO had been effected. Here, we identify the potential source of this NO. Treatment of leaves with antimycin A (AA, Q_i‐site inhibitor of Complex III) increased NO amount more than treatment with myxothiazol (Myxo, Q_o‐site inhibitor) despite both being equally effective at inhibiting respiration. Comparison of nitrate‐grown wild‐type with AOX knockdown and overexpression plants showed a negative correlation between AOX amount and NO amount following AA. Further, Myxo fully negated the ability of AA to increase NO amount. With ammonium‐grown plants, neither AA nor Myxo strongly increased NO amount in any plant line. When these leaves were supplied with nitrite alongside the AA or Myxo, then the inhibitor effects across lines mirrored that of nitrate‐grown plants. Hence the ETC, likely the Q‐cycle of Complex III generates NO from nitrite, and AOX reduces this activity by acting as a non‐energy‐conserving electron sink upstream of Complex III.     

Optimization and scale up of itaconic acid production from potato starch waste in stirred tank bioreactor

Present study used Aspergillus terreus strain C1 isolated from mangrove soil for itaconic acid (IA) production from potato starch waste. Fermentation parameters were optimized by classical one factor approach and statistical experimental designs, such as Plackett-Burman and response surface designs. Anionic deionization of potato waste was found to be a very effective, economic, and easy way of improving IA production. The increase in IA production by deionization was found to correlate with removal of phosphate. In our knowledge, this is the first report on application of deionization of potato waste to enhance IA production. Other parameters like inoculum development conditions, pH, presence of peptone and certain salts in the medium also significantly affected IA production. IA production by strain C1 increased 143-fold during optimization when compared with the starting condition. The optimized IA level (35.75 g/L) was very close to the maximum production predicted by RSM (38.88 g/L). Bench scale production of IA was further optimized in 3-L stirred tank reactor by varying parameters like agitation and aeration rate. The maximum IA production of 29.69 g/L was obtained under the agitation speed of 200 rpm and aeration rate of 0.25 vvm. To the best of our knowledge, it is the first report on IA production from potato starch waste at bioreactor level. © 2019 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2774, 2019.     

Micropropagation,berberine content and antitumor activity of Jeffersonia dubia (Maxim.) Benth et Hook

An efficient micropropagation protocol was developed for Jeffersonia dubia using sucker explants. High frequency of multiple shoot formation was induced when the sucker explants were cultured on Chu’s (N6) medium with different concentrations of thidiazuron (TDZ) plus 0.54 µM α-naphthaleneacetic acid (NAA). The maximum frequency of shoot formation (96.2 %) was obtained on N6 medium with 2.27 µM TDZ plus 0.54 µM NAA. The highest mean number of shoots per explant (13.6) was obtained in temporary immersion system using an immersion frequency of 30 s every 30 min. The highest frequency of rooting (100 %), number of roots per shoot (5.8), and root length (6.3) was observed in half-strength N6 medium supplemented with 2.69 µM NAA. The regenerated plantlets (30 days old) were successfully acclimatized in the greenhouse with 98 % survival rate. The berberine content and cytotoxicity were higher in in vitro-developed calli and shoots than in leaves of field-grown plants. The greatest content of berberine was found in shoots (1381 μg g⁻¹) followed by calli (1092 μg g⁻¹) and leaves of field-grown plants (92 μg g⁻¹). At 1000 μg mL⁻¹ concentration, growth inhibition rate of berberine, callus, shoot, and leaf (in vivo) extracts were 68.4, 57.1, 54.2, and 17.7 %, respectively.

     

Two-step biodiesel production from Calophyllum inophyllum oil: Optimization of modified β-zeolite catalyzed pre-treatment

In this study, a two-step process was developed to produce biodiesel from Calophyllum inophyllum oil. Pre-treatment with phosphoric acid modified β-zeolite in acid catalyzed esterification process preceded by transesterification which was done using conventional alkali catalyst potassium hydroxide (KOH). The objective of this study is to investigate the relationship between the reaction temperatures, reaction time and methanol to oil molar ratio in the pre-treatment step. Central Composite Design (CCD) and Response Surface Methodology (RSM) were utilized to determine the best operating condition for the pre-treatment step. Biodiesel produced by this process was tested for its fuel properties.     

Itaconic Acid Production by Filamentous Fungi in Starch-Rich Industrial Residues

Several fungi and starch-rich industrial residues were screened for itaconic acid (IA) production. Out of 15 strains, only three fungal strains were found to produce IA, which was confirmed by HPLC and GC–MS analysis. These strains were identified as Aspergillus terreus strains C1 and C2, and Ustilago maydis strain C3 by sequencing of 18S rRNA gene and internal transcribed spacer regions. Cis-aconitate decarboxylase (cad) gene, which encodes a key enzyme in IA production in A. terreus, was characterized from strains C1 and C2. C1 and C2 cad gene sequences showed about 96% similarity to the only available GenBank sequence of A. terreus cad gene. 3-D structure and cis-aconitic acid binding pocket of Cad enzyme were predicted by structural modeling. Rice, corn and potato starch wastes were screened for IA production. These materials were enzymatically hydrolyzed under experimentally optimized conditions resulting in the highest glucose production of 230 mg/mL from 20% potato waste. On comparing the production potential of selected strains with different wastes, the best IA production was achieved with strain C1 (255.7 mg/L) using potato waste. Elemental composition as well as batch-to-batch variation in waste substrates were analyzed. The difference in IA production from two different batches of potato waste was found to inversely correlate with their phosphorus content, which indicated that A. terreus produced IA under phosphate limiting condition. The potato waste hydrolysate was deionized to remove inhibitory ions like phosphate, resulting in improved IA production of 4.1 g/L by C1 strain, which is commercially competitive.     

Retinoic Acid Mediates Regulation of Network Formation by COUP-TFII and VE-Cadherin Expression by TGFβ Receptor Kinase in Breast Cancer Cells

Tumor development, growth, and metastasis depend on the provision of an adequate vascular supply. This can be due to regulated angiogenesis, recruitment of circulating endothelial progenitors, and/or vascular transdifferentiation. Our previous studies showed that retinoic acid (RA) treatment converts a subset of breast cancer cells into cells with significant endothelial genotypic and phenotypic elements including marked induction of VE-cadherin, which was responsible for some but not all morphological changes. The present study demonstrates that of the endothelial-related genes induced by RA treatment, only a few were affected by knockdown of VE-cadherin, ruling it out as a regulator of the RA-induced endothelial genotypic switch. In contrast, knockdown of the RA-induced gene COUP-TFII prevented the formation of networks in Matrigel but had no effect on VE-cadherin induction or cell fusion. Two pan-kinase inhibitors markedly blocked RA-induced VE-cadherin expression and cell fusion. However, RA treatment resulted in a marked and broad reduction in tyrosine kinase activity. Several genes in the TGFβ signaling pathway were induced by RA, and specific inhibition of the TGFβ type I receptor blocked both RA-induced VE-cadherin expression and cell fusion. Together these data indicate a role for the TGFβ pathway and COUP-TFII in mediating the endothelial transdifferentiating properties of RA.