Identification of a potential fungal species by 18S rDNA for ligninases production

Fungal species for ligninases production was investigated by 18S ribosomal DNA sequence analysis. Two primer sets were chosen to amplify a major part of the 18S rDNA, which resulted in intense PCR product of approximately 550–820 bp in size per sample. The results suggest that the 18S rDNA-based approach is a useful tool for identification of unknown potential fungal species for ligninases production. The isolated fungal species produces mainly manganese peroxidase (MnP). The enzyme oxidized a variety of the usual MnP substrates, including lignin related polyphenols. Time course studies showed that maximum production of ligninolytic enzymes MnP (64 IU L^?1), lignin peroxidase (26.35 IU L^?1), and laccase (5.44 IU L^?1), respectively, were achieved after 10 days of cultivation under optimum conditions. Furthermore, the biological decolorization of Remazol Brilliant Blue R dye following 10 days of cultivation was 94 %. NCBI BLAST was used to search for closest matched sequences in the GenBank database and based on sequence homology the first BLAST hit was Dothioraceae sp. LM572 with accession number EF060858.1.     

Enzymatic deinking of laser printed office waste papers: some governing parameters on deinking efficiency

The protocol for the enzymatic deinking of laser printed waste papers on a laboratory scale using cellulase (C) and hemicellulase (H) of Aspergillus niger (Amano) was developed as an effective method for paper recycling. A maximum deinking efficiency of almost 73% by the enzyme combination of C:H was obtained using the deinking conditions of pulping consistency of 1.0% (w/v) with the pulping time of 1.0min, temperature of 50 degrees C, pH=3.5, agitation rate of 60rpm, pulp concentration of 4% (w/v), concentration of each enzyme of 2.5U/g air dried pulp and the enzyme ratio of 1:1. The deinking efficiency was further enhanced to 95% using the optimized flotation system consisting of pH=6.0, Tween 80 of concentration 0.5% (w/w), working air flow rate of 10.0L/min and temperature of 45 degrees C. The deinked papers were found to exhibit properties comparable to the commercial papers suggesting the effectiveness of the enzymatic process developed.     

Highly thermo–halo–alkali-stable β-1,4-endoxylanase from a novel polyextremophilic strain of Bacillus halodurans

A novel bacterial isolate, capable of producing extracellular highly thermostable, halo-alkali-stable and cellulase-free xylanase, was isolated from soil and identified as Bacillus halodurans TSPV1 by polyphasic approach. The Plackett–Burman design identified wheat bran, lactose, tryptone and NaCl as the factors that significantly affect xylanase production, and thus, these were optimized by response surface methodology. The data analysis suggested that optimum levels of wheat bran (15–20 g L^?1), lactose (1.0–1.5 g L^?1), tryptone (2–2.5 g L^?1) and NaCl (7.0–8.0 g L^?1) support 6.75-fold higher xylanase production than that in the un-optimized medium. The xylanase is optimally active at 90 °C and pH 10, and stable for 4 h at 90 °C (T _1/2 60 h) over a broad range of NaCl concentrations (0–29 %). This is the first report on the isolation of polyextremophilic B. halodurans strain that produces thermo-halo-alkali-stable xylanase in submerged fermentation. This enzyme efficiently saccharifies agro residues like wheat bran and corncobs. Fifty-six percent of hemicellulose of wheat bran could be hydrolyzed by xylanase (100 U g^?1 substrate) along with cellulase (22 U FPase and 50 U CMCase g^?1). The xylanase, being thermo-alkali stable and cellulase free, can find applications in pre-bleaching of paper pulps and hydrolysis of xylan in agricultural residues.     

Combined Enzymatic and Physical Deinking Methodology for Efficient Eco-Friendly Recycling of Old Newsprint

Background

The development in the deinking process has made recycled fiber a major part of the raw material for pulp and paper industry. Enzymes have revolutionized the deinking process obtaining brightness levels surpassing conventional deinking processes. This study explores the deinking efficiencies of bacterial alkalophilic laccase (L) and xylanase (X) enzymes along with physical deinking methods of microwaving (MW) and sonication (S) for recycling of old newsprint (ONP).

Methods and Results

The operational parameters viz. enzyme dose, pH and treatment time for X and L deinking were optimized statistically using Response Surface Methodology. Laccase did not require any mediator supplementation for deinking. Deinking of ONP pulp with a combination of xylanase and laccase enzymes was investigated, and fiber surface composition and morphological changes were studied using X-ray diffraction, fourier transform infrared spectroscopy and scanning electron microscopy. Compared to the pulp deinked with xylanase (47.9%) or laccase (62.2%) individually, the percentage reduction of effective residual ink concentration (ERIC) was higher for the combined xylanase/laccase-deinked pulp (65.8%). An increase in brightness (21.6%), breaking length (16.5%), burst factor (4.2%) tear factor (6.9%), viscosity (13%) and cellulose crystallinity (10.3%) along with decrease in kappa number (22%) and chemical consumption (50%) were also observed. Surface appeared more fibrillar along with changes in surface functional groups. A combination of physical and enzymatic processes (S-MW-XL) for deinking further improved brightness (28.8%) and decreased ERIC (73.9%) substantially.

Conclusion

This is the first report on deinking of ONP with laccase without any mediator supplementation. XL pretreatment resulted in marked improvement in paper quality and a new sequence being reported for deinking (S-MW-XL) will contribute further in decreasing chemical consumption and making the process commercially feasible.     

Characterization of a novel asparaginase from soil metagenomic libraries generated from forest soil

Objectives

To screen soil metagenomic libraries for novel enzymes with enhanced activities.

Results

To screen soil metagenomic libraries for novel enzymes with enhanced activities. A novel l-asparaginase was identified from forest soil metagenome and its characteristics were studied. The purified protein had a specific activity of 696 IU mg^?1 and optimum activity at pH 7 and 35 °C. Enhanced enzyme activities were observed in the presence of Mg²⁺, Ca²⁺ and K⁺. The K_m value, 2 mM, and enzyme specificity constant 7.7 mM^?1s^?1 indicated that the recombinant enzyme has good substrate affinity to l-asparagine compared with commercially-available Escherichia coli asparaginase. The IC₅₀ value of 0.78 µg ml^?1 (0.47 IU ml^?1) was observed with HL60 cell line and 0.39 µg ml^?1(0.23 IU ml^?1) with MOLT-3 and MOLT-4 cell lines, which is better than that of commercially-available drugs.

Conclusion

The soil metagenome derived l-asparaginase with enhanced activities could be a potential candidate to develop as a drug in Acute Lymphoblastic Leukemia (ALL) therapy.

     

Hyper production of cellulase-free xylanase by <Emphasis Type="Italic">Thermomyces lanuginosus</Emphasis> SSBP on bagasse pulp and its application in biobleaching

A cellulase-free xylanase production by Thermomyces lanuginosus SSBP using bagasse pulp was examined under submerged (SmC) and solid-state cultivation (SSC). Higher level of xylanase activity (19,320 ± 37 U g⁻¹ dried carbon source) was obtained in SSC cultures than in SmC (1,772 ± 15 U g⁻¹ dried carbon source) after 120 h with 10% inoculum. The biobleaching efficacy of crude xylanase was tested on bagasse pulp, and the maximum brightness of 46.1 ± 0.06% was observed with 50 U of crude xylanase per gram of pulp, which was 3.8 points higher than the brightness of untreated samples. Reducing sugars (26 ± 0.1 mg g⁻¹) and UV-absorbing lignin-derived compounds in the pulp filtrates were observed as maximum in 50 U of crude xylanase-treated samples. T. lanuginosus SSBP has potential applications due to its high productivity of xylanase and its efficiency in pulp bleaching.     

Process optimization of xylanase production using cheap solid substrate by Trichoderma reesei SAF3 and study on the alteration of behavioral properties of enzyme obtained from SSF and SmF

This study aimed to assess the variability in respect of titer and properties of xylanase from Trichoderma reesei SAF3 under both solid-state and submerged fermentation. SSF was initially optimized with different agro-residues and among them wheat bran was found to be the best substrate that favored maximum xylanase production of 219 U (gws)^?1 at 96 h of incubation. The mycelial stage of the fungi and intracellular accumulation of Ca⁺⁺ and Mg⁺⁺ induced maximum enzyme synthesis. Inoculum level of 10 × 10⁶ spores 5 g^?1 of dry solid substrate and water activity of 0.6 were found to be optimum for xylanase production under SSF. Further optimization was made using a Box-Behnken design under response surface methodology. The optimal cultivation conditions predicted from canonical analysis of this model were incubation time (A) = 96–99 h, inoculum concentration (B) = 10 × 10⁶ spores 5 g^?1 of dry substrate, solid substrate concentration (C) = 10–12 g flask^?1, initial moisture level (D) = 10 mL flask^?1 (equivalent to a _w  = 0.55) and the level of xylanase was 299.7 U (gws)^?1. Subsequent verification of these levels agreed (97 % similar) with model predictions. Maximum amount of xylanase was recovered with water (6:1, v/w) and under shaking condition (125 rpm). Purified xylanase from SSF showed better stability in salt and pH, was catalytically and thermodynamically more efficient over enzyme from SmF, though molecular weight of both enzymes was identical (53.8 kDa).     

Biobleaching of banana fibre pulp using <Emphasis Type="Italic">Bacillus subtilis</Emphasis> C O1 xylanase produced from wheat bran under solid-state cultivation

A cellulase-free xylanase produced by Bacillus subtilis C 01 from wheat bran under solid-state cultivation was tested for its efficacy in biobleaching of raw banana fibre and banana pulp obtained through a mechanical pulping process. Banana pulp samples treated with crude xylanase (450 nkat g⁻¹ pulp) resulted in a 19.6% increase in the brightness as compared to untreated pulp. The presence of chromophores, hydrophobic compounds and an increased reducing sugar (10.79 mg g⁻¹ pulp) quantity in the bleached solution after enzymatic treatment indicated the removal of materials that were absorbed at 237 nm from the banana pulp.     

Determination of resistance to low temperatures of winter buds on lateral shoot present in Karaerik (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L.) grape cultivar

This study aims to determine the low temperature resistance of dormant buds at nodes with or without lateral shoots of Karaerik grape cultivar, and explain the relationship between the resistance and biochemical parameters in this grape cultivar. In this study, the mean values of high temperature exotherms (HTEs), low temperature exotherms (LTEs), water, reducing sugar, total soluble protein contents and antioxidant enzyme activities of dormant buds taken from nodes with or without lateral shoots were determined. The experiment has been found that buds in nodes with lateral shoots showed HTE and LTE at higher temperatures (HTE average ?6.7 °C and LTE average ?8.3 °C) than buds (HTE average ?7.9 °C and LTE average ?11.5 °C) in nodes without lateral shoots; therefore, buds in nodes with lateral shoots had less tolerance to low temperature. Additionally, lower sugar (average 41.05 mg g^?1), protein (average 1.61 mg g^?1), superoxide dismutase (average 425.27 EU g^?1 tissue), peroxidase (average 2516.1 EU g^?1 tissue) and polyphenol oxidase (average 7283.1 EU g^?1 tissue) were determined for buds taken from nodes with lateral shoots. Due to the fact that dormant buds taken from nodes with lateral shoots decreased the resistance to low temperatures, this research suggests that these lateral shoots should be excised with the summer pruning at the regions, where low temperatures caused the damages.     

Characterization of a novel xylanase from Armillaria gemina and its immobilization onto SiO2 nanoparticles

Enhanced catalytic activities of different lignocellulases were obtained from Armillaria gemina under statistically optimized parameters using a jar fermenter. This strain showed maximum xylanase, endoglucanase, cellobiohydrolase, and β-glucosidase activities of 1,270, 146, 34, and 15 U mL^?1, respectively. Purified A. gemina xylanase (AgXyl) has the highest catalytic efficiency (k _cat/K _m?=?1,440 mg?mL^?1?s^?1) ever reported for any fungal xylanase, highlighting the significance of the current study. We covalently immobilized the crude xylanase preparation onto functionalized silicon oxide nanoparticles, achieving 117 % immobilization efficiency. Further immobilization caused a shift in the optimal pH and temperature, along with a fourfold improvement in the half-life of crude AgXyl. Immobilized AgXyl gave 37.8 % higher production of xylooligosaccharides compared to free enzyme. After 17 cycles, the immobilized enzyme retained 92 % of the original activity, demonstrating its potential for the synthesis of xylooligosaccharides in industrial applications.     

Effect of elicitation and precursor feeding on accumulation of 20-hydroxyecdysone in <Emphasis Type="Italic">Achyranthes aspera</Emphasis> Linn. cell suspension cultures

20-Hydroxyecdysone is one of the most common ecdysteroids in plants with potential therapeutic applications. In this study, cell suspension cultures of Achyranthes aspera were raised in shake flasks to investigate the production of 20-hydroxyecdysone. The quantification and characterization of 20-hydroxyecdysone in the cultures were done by High performance liquid chromatography (HPLC) and Liquid Chromatography-quadrupole time-of- flight mass spectrometry (LC-Q-TOF) analyses. For raising the suspension, calli initiated from in vitro grown leaf explants were cultured in liquid Murashige and Skoog (MS) medium augmented with combinations of 2, 4-dichlorophenoxyacetic acid (1 mg L^?1) and α-naphthaleneacetic acid (1 mg L^?1). Maximum growth index of the cell suspension was 9.9, which was achieved during 20th day of culture (final phase of exponential growth). At this stage, the biomass accumulated was 1.09 ± 0.09 g dry weight (DW) and the 20-hydroxyecdysone concentration was 0.24 mg g^?1 DW. Eliciting the cultures with 0.6 mM Methyl jasmonate for 6 days; enhanced the production of 20-hydroxyecdysone production to 0.35 mg g^?1 DW. By augmenting the cultures with the precursors namely cholesterol (10 mg L^?1) and 7-dehydrocholesterol (10 mg L^?1), production of 20-hydroxyecdysone was boosted to 0.31 mg g^?1 DW and 0.28 mg g^?1 DW respectively.     

Manipulation of culture strategies to enhance capsaicin biosynthesis in suspension and immobilized cell cultures of Capsicum chinense Jacq. cv. Naga King Chili

Manipulation of culture strategies was adopted to study the influence of nutrient stress, pH stress and precursor feeding on the biosynthesis of capsaicin in suspension and immobilized cell cultures of C. chinense. Cells cultured in the absence of one of the four nutrients (ammonium and potassium nitrate for nitrate and potassium stress, potassium dihydrogen orthophosphate for phosphorus stress, and sucrose for sugar stress) influenced the accumulation of capsaicin. Among the stress factors studied, nitrate stress showed maximal capsaicin production on day 20 (505.9 ± 2.8 μg g^?1 f.wt) in immobilized cell, whereas in suspension cultures the maximum accumulation (345.5 ± 2.9 μg g^?1 f.wt) was obtained on day 10. Different pH affected capsaicin accumulation; enhanced accumulation of capsaicin (261.6 ± 3.4 μg g^?1 f.wt) was observed in suspension cultures at pH 6 on day 15, whereas in case of immobilized cultures the highest capsaicin content (433.3 ± 3.3 μg g^?1 f.wt) was obtained at pH 5 on day 10. Addition of capsaicin precursors and intermediates significantly enhanced the biosynthesis of capsaicin, incorporation of vanillin at 100 μM in both suspension and immobilized cell cultures resulted in maximum capsaicin content with 499.1 ± 5.5 μg g^?1 f.wt on day 20 and 1,315.3 ± 10 μg g^?1 f.wt on day 10, respectively. Among the different culture strategies adopted to enhance capsaicin biosynthesis in cell cultures of C. chinense, cells fed with vanillin resulted in the maximum capsaicin accumulation. The rate of capsaicin production was significantly higher in immobilized cells as compared to freely suspended cells.     

Enzymatic deinking of various types of waste paper: Efficiency and characteristics

The aim of the present study was to characterize the enzymatic deinking of various types of waste paper. Studies on the optimization of enzymatic deinking have been performed previously using commercially available enzyme preparations containing cellulase and hemicellulase. The enzymatic deinking of different types of waste paper demonstrated a high efficiency of 86.6% on laser-printed paper, but a low deinking efficiency of 12.9% was obtained with newspaper. All enzymatic treatments significantly improved the drainage rate of the deinked waste paper. Enzymatic deinking increased the tensile index of magazine paper but reduced the tensile index of bubble jet-printed paper, photocopy paper and newspaper. Enzymatic hydrolysis caused a 21.1% reduction in the tear index for bubble jet-printed paper, but a 3.1% increase in the tear index was obtained for laser-printed paper relative to respective blank. In addition, enzymatic hydrolysis increased the burst index by 4.7% relative to blank for laser-printed paper. However, photocopy paper showed the highest reduction (8.3%) in the burst index relative to blank. Taken together, these results suggest that enzymatic hydrolysis is both advantageous and detrimental to the mechanical properties of deinked paper. Thus, the proper regulation of enzymatic hydrolysis is crucial to improve the quality of recycled paper.     

Fiber surface characterization of old newsprint pulp deinked by combining hemicellulase with laccase-mediator system

Deinking of old newsprint (ONP) by combining hemicellulase with laccase-mediator system (LMS) was investigated, and surface chemical composition and fiber morphology changes during the deinking process were studied by electron spectroscopy for chemical analysis (ESCA), contact angle (CA), attenuated total reflectance fourier transform infrared spectrometry (ATR-FTIR), fiber quality analyzer (FQA), and environmental scanning electronic microscopy (ESEM). Results showed that, compared to the pulp deinked with hemicellulase or LMS individually, effective residual ink concentration (ERIC) was lower for the hemicellulase/LMS-deinked pulp. This indicated that there is a synergistic deinking effect between hemicellulase and LMS. It was found that O/C ratio of the fiber surface increased and the surface coverage of lignin decreased during the hemicellulase/LMS deinking process. The contact angle of the hemicellulase/LMS-deinked pulp was lower than that of pulps deinked with each individual enzyme. ESEM observations showed that more fibrils appeared on the fiber surface due to synergistic treatment.     

Production of thermo-alkali-stable xylanase by a novel polyextremophilic Bacillus halodurans TSEV1 in cane molasses medium and its applicability in making whole wheat bread

A high titre of thermo-alkali-stable xylanase was attained in cane molasses medium. When the culture variables for endoxylanase production were optimized [cane molasses 7 %, soluble alkaline extract of wheat bran (SAE-WB) 37 % and ammonium chloride 0.30 %], a 4.5-fold enhancement in xylanase production (69 U ml^?1) was achieved as compared to that in the unoptimized medium (15 U ml^?1). The enzyme titre attained in shake flasks could be sustained in a 7-l laboratory bioreactor. An activity band corresponding to 40 kDa was visualized on SDS-PAGE zymogram analysis. The enzyme has broad range of pH and temperature for activity with optima at 9.0 and 80 °C, and stable between pH 4.0 and 11.0 with 85 % retention of activity. It has T _1/2 of 40 and 15 min at 70 and 80 °C. The enzyme is halotolerant since it displays activity in the presence of salt up to 15 %, and remains 100 % active in the absence of salt. The supplementation of whole wheat dough with xylanase improves antistaling property, reducing sugar content, bread volume with prebiotic xylooligosaccharides in bread. This is the first report on xylanase production in cane molasses medium with SAE-WB as the inducer and its applicability in whole wheat bread making that improves human health.     

Nitrogen Translocation to Fresh Litter in Northern Hardwood Forest

Nitrogen immobilization in fresh litter represents a significant N flux in forest ecosystems, and changes in this process resulting from atmospheric N deposition could have important implications for ecosystem responses. We conducted two leaf decay experiments, using ¹⁵N-labeled sugar maple leaf litter, to quantify N transport from old litter and soil to fresh litter during early stages of decomposition, and we examined the influence of litter N concentration and soil N availability on upward N transfer in a northern hardwood forest. After one year of decay, the average N transfer from soil to fresh litter (2.63 mg N g^?1 litter) was much higher than the N transfer from older litter (1- to 2-years-old) to fresh litter (0.37 mg N g^?1 litter). We calculated the ratio of annual N transfer per unit of excess ¹⁵N pool for these two N sources. The ratio was not significantly different between old litter and soil, suggesting that fungi utilize N in the old litter and mineral soil pools for transport to decaying fresh litter with similar efficiency. Initial litter N concentration had a significant effect on upward N flux into decaying leaf litter, whereas no effect of soil N fertilization was observed. Reduction in the flux from soil to fresh litter owing to anthropogenic N inputs probably contributes significantly to changing soil N dynamics. Future work is needed on fungal N acquisition and transport as well as the fungal taxa involved in this process and their responses to changing environments.     

The carbohydrate-binding module of Fragaria × ananassa expansin 2 (CBM-FaExp2) binds to cell wall polysaccharides and decreases cell wall enzyme activities “in vitro”

A putative carbohydrate binding module (CBM) from strawberry (Fragaria × ananassa Duch.) expansin 2 (CBM-FaExp2) was cloned and the encoding protein was over-expressed in Escherichia coli and purified in order to evaluate its capacity to bind different cell wall polysaccharides “in vitro”. The protein CBM-FaExp2 bound to microcrystalline cellulose, xylan and pectin with different affinities (K_ad = 33.6 ± 0.44 mL g^?1, K_ad = 11.37 ± 0.87 mL g^?1, K_ad = 10.4 ± 0.19 mL g^?1, respectively). According to “in vitro” enzyme assays, this CBM is able to decrease the activity of cell wall degrading enzymes such as polygalacturonase, endo-glucanase, pectinase and xylanase, probably because the binding of CBM-FaExp2 to the different substrates interferes with enzyme activity. The results suggest that expansins would bind not only cellulose but also a wide range of cell wall polymers.     

Feeding elicitors and precursors enhance colchicine accumulation in morphogenic cultures of <Emphasis Type="Italic">Gloriosa superba</Emphasis> L.

Morphogenic cultures of Gloriosa superba were initiated on Murashige and Skoog’s medium fortified with 2 mg L^?1 2,4-dichlorophenoxyacetic acid (2,4-D), 0.5 mg L^?1 naphthaleneacetic acid (NAA), 4% sucrose and 0.1% activated charcoal. To enhance the content of the alkaloid colchicine, morphogenic cultures were treated with different concentrations of abiotic elicitors like signalling compounds, metals, biotic elicitors, precursors and a combination of elicitors. Signalling molecules like acetyl salicylic acid (ASA) and sodium nitroprusside improved the production of colchicine. Abiotic elicitors have markedly (p?≤?0.05 or ≤?0.01) enhanced the colchicine content either at lower or higher concentrations. Among the metals, the highest amount of 11.67 mg of colchicine g^?1 dry wt was noticed at 60 mM rubidium chloride, followed by 60 mM NaCl (11.18 mg g^?1). Contrarily, in the presence of biotic elicitors such as Fusarium oxysporum, Alternaria solani, and Saccharomyces cerevisiae, colchicine content ranged only between 2 and 5.32 mg g^?1, but Bacillus subtilis repressed it. Among the aromatic amino acids, phenylalanine at 500 mg L^?1 influenced the highest accumulation of 19.48 mg g^?1 dry tissue, followed by tryptophan (12.47 mg g^?1), and tyrosine (9.87 mg g^?1), a direct precursor of colchicine biosynthesis, while intact tubers and leaves contained 4.65 and 4.16 mg of colchicine g^?1 dry tissue respectively. A combination of 10 µM AlCl₃ and 50 µM salicylic acid (SA) registered 17.34 mg g^?1 followed by 16.24 mg g^?1 tissue in presence of 1 µM HgCl₂ and 50 µM SA. The results suggest that the elicitor-stimulated colchicine accumulation was a stress response and can be exploited further for commercial production.     

Light-induced production of antidepressant compounds in etiolated shoot cultures of Hypericum hookerianum Wight & Arn. (Hypericaceae)

Hypericum hookerianum is a lesser known ethnomedicinal plant having wound healing, antitumor and anti-HSV-1 properties. Isolated nodes of in vitro shoots sub-cultured in the dark for 4 weeks on half strength Murashige and Skoog medium solidified with Gelzan (1.5 g l^?1), and supplemented with 2.325 μM kinetin produced 8.0 ± 0.40 etiolated shoots of 5.0 ± 0.62 cm length at 74 % efficiency versus 9.2 ± 0.6 healthy shoots of 4.4 ± 0.5 cm obtained from nodes in light at 96 % efficiency. Low concentrations of hypericin were found in wild plant [0.35 ± 0.09 mg g^?1 dry weight (DW)] and control green shoot cultures (0.91 ± 0.03 mg g^?1 DW). Etiolated shoots exposed to a 12 h photoperiod (50 μmol m^?2 s^?1) through 1–25 days turned red incrementally due to synthesis and accumulation of 0.1–3.83 mg g^?1 DW hypericin in sub-epidermal cortical cells of the stem and varied shaped cells of the distorted mesophyll. Flavonoid and anthocyanin concentrations of the etiolated shoots subjected to the 12 h photoperiod were 3–5 fold higher than the control shoot cultures while total chlorophylls [1.97 ± 0.05 mg g^?1 fresh weight (FW)] of the light exposed shoots were significantly less compared to the control (2.86 ± 0.18 mg g^?1 FW) and natural plant (6.82 ± 0.29 mg g^?1 FW). HPLC analysis of shoot extracts revealed the presence of 0.14 ± 0.03, 0.16 ± 0.02 and 1.45 ± 0.16 mg g^?1 DW hyperforin in wild plant, control shoot cultures and etiolated shoot cultures illuminated for 25 days, respectively. Despite a reasonable presence in etiolated shoots (0.61 ± 0.15 g^?1 FW), total phenols did not increase significantly during illumination. The results indicate light induced synthesis of anti-depressant phenolic derivatives (hypericin, hyperforin and flavonoids) in etiolated shoot cultures of H. hookerianum.     

Xylan-hydrolyzing thermotolerant <Emphasis Type="Italic">Candida tropicalis</Emphasis> HNMA-1 for bioethanol production from sugarcane bagasse hydrolysate

Sugarcane bagasse is one of the low-cost substrates used for bioethanol production. In order to solubilize sugars in hemicelluloses like xylan, a new thermotolerant isolate of Candida tropicalis HNMA-1 with xylan-hydrolyzing ability was identified and characterized. The strain showed relative tolerance to high temperature. Our results demonstrated 0.211 IU ml^?1 xylanase activity at 40 °C compared to 0.236 IU ml^?1 at 30 °C. The effect of high temperature on the growth and fermentation of xylose and sugarcane bagasse hydrolysate were also investigated. In both xylose or hydrolysate medium, increased growth was recorded at 40 °C. Meanwhile, the efficiency of ethanol fermentation was adversely affected by temperature since yields of 0.088 g g^?1 and 0.076 g g^?1 in the xylose medium, in addition to 0.090 g g^?1 and 0.078 g g^?1 in the hydrolysate medium were noticed at 30 °C and 40 °C, respectively. Inhibitory compounds in the hydrolysate medium demonstrated negative effects on fermentation and productivity, with maximum ethanol concentration attained after 48 h in the hydrolysate, as opposed to 24 h in the xylose medium. Our data show that the newly thermotolerant isolate, C. tropicalis HNMA-1, is able to efficiently ferment xylose and hydrolysate, and also has the capacity for application in ethanol production from hemicellulosic sources.