Effect of soybean oil and Tween 80 on the production of botryosphaeran by Botryosphaeria rhodina MAMB-05

The addition of soybean oil and Tween 80 was evaluated with the objective of increasing the production of botryosphaeran, an exopolysaccharide (EPS) of the (1 → 3;1 → 6)-β-d-glucan type produced by the fungus Botryosphaeria rhodina MAMB-05. Factorial design and analysis by response surface methodology was developed to select the main factors that would affect and enhance EPS production. The optimized culture conditions were: 40 g l⁻¹ glucose with 10 ml l⁻¹ soybean oil, and 4.5 ml l⁻¹ Tween 80, during 72 h cultivation at 28 °C (180 rpm) and initial pH 5.7. The predicted result for botryosphaeran production was 8.22 ± 1.36 g l⁻¹, and compared with the experimental value of 7.74 ± 0.13 g l⁻¹. Partial characterization of the botryosphaeran produced under the optimized conditions showed one type of polysaccharide with β-glycosidic linkages containing glucose as monosaccharide.     

Effects of feed sugar concentration on continuous ethanol fermentation of cheese whey powder solution (CWP)

Cheese whey powder (CWP) solution with different CWP or sugar concentrations was fermented to ethanol in a continuous fermenter using pure culture of Kluyveromyces marxianus (DSMZ 7239). Sugar concentration of the feed CWP solution varied between 55 and 200 g l⁻¹ while the hydraulic residence time (HRT) was kept constant at 54 h. Ethanol formation, sugar utilization and biomass formation were investigated as functions of the feed sugar concentration. Percent sugar utilization and biomass concentrations decreased and the effluent sugar concentration increased with increasing feed sugar concentrations especially for the feed sugar contents above 100 g l⁻¹. Ethanol concentration and productivity (DP) increased with increasing feed sugar up to 100 g l⁻¹ and then decreased with further increases in the feed sugar content. The highest ethanol concentration (3.7%, v v⁻¹) and productivity (0.54 gE l⁻¹ h⁻¹) were obtained with the feed sugar content of 100 g l⁻¹ or 125 g l⁻¹. The ethanol yield coefficient (Y_P/S) was also maximum (0.49 gE gS⁻¹) when the feed sugar was between 100 and 125 g l⁻¹. The growth yield coefficient (Y_X/S) decreased steadily from 0.123 to 0.063 gX gS⁻¹ when the feed sugar increased from 55 to 200 g l⁻¹ due to adverse effects of high sugar contents on yeast growth. The optimal feed sugar concentration maximizing the ethanol productivity and sugar utilization was between 100 and 125 g l⁻¹ under the specified experimental conditions.     

Signal transducer and oxidative stress mediated modulation of phenylpropanoid pathway to enhance rosmarinic acid biosynthesis in fungi elicited whole plant culture of Solenostemon scutellarioides

This study aimed to improve rosmarinic acid (RA) production in the whole plant culture of Solenostemon scutellarioides through elicitation with phytopathogenic fungi. Amongst selected fungi, Aternaria alternata caused significant elevation (p < 0.05–0.01) in RA accumulation (∼1.3–1.6-fold) between 25 and 100 μg l⁻¹. However, elicitation at the dose of 50 μg l⁻¹ has been found to be most effective and intracellular RA content reached almost ∼1.6-fold (p < 0.01) higher in day 7. Therefore, A. alternata (50 μg l⁻¹) was selected for mechanism evaluation. A significant elevation of intercellular jasmonic acid was observed up to day 6 after elicitation with A. alternata (50 μg l⁻¹). A significant increase in tissue H₂O₂ and lipid peroxidation coupled with depletion of antioxidant enzymes superoxide dismutase and catalase indicated augmented oxidative stress associated with biotic interaction. Preceding the elicitor-induced RA accumulation, a notable alteration in the specific activities of biosynthetic enzymes namely PAL and TAT was recorded, while, no significant change in the activities of RAS was observed. HPPR activity was slightly improved in elicited plant. Therefore, it could be concluded that A. alternata elicited the biosynthesis of rosmarinic acid via signal transduction through jasmonic acid coupled with elicitor induced oxidative stress and associated mechanism.     

Isolation of hexavalent chromium resistant bacteria from industrial saline effluents and their ability of bioaccumulation

The mixed cultures has been isolated from industrial saline wastewater contaminated with chromium(VI), using enrichment in the presence of 50 mg l⁻¹ chromium(VI) and 4% (w/v) NaCl at pH 8. In this study, the molasses (M) medium was selected a suitable medium for the effective chromium bioaccumulation by the mixed cultures. Eleven pure isolates obtained from mixed cultures and some of them showed high bioaccumulation in the M media containing about 100 mg l⁻¹ chromium(VI) and 4% NaCl. The strain 8 (99.3%) and 10 (99.1%) were able to bioaccumulate more efficient than the mixed culture (98.9%) in this media. But the highest specific Cr uptake was obtained by the mixed cultures followed by strain 8 and 10 with 56.71, 33.14 and 21.7 mg g⁻¹, respectively. Bioaccumulation of chromium(VI) ions by the strain 8 growing in the media with chromium(VI) and NaCl was studied in a batch system as a function of initial chromium(VI) (86.6–547.6 mg l⁻¹) and NaCl (0, 2, 4, 6% w/v) concentrations. During all the experiments, the uptake yield of the strain 8 was highly affected from NaCl concentrations in the medium at high initial chromium(VI) concentrations. But at low chromium(VI) concentration, strain 8 was not affected from NaCl concentrations in the medium. The maximum uptake yield were obtained in the M media with 2% NaCl as 98.8% for 110.0 mg l⁻¹, 98.6% for 217.1 mg l⁻¹, 98.6% for 381.7 mg l⁻¹ and 98.2% for 547.6 mg l⁻¹ initial chromium(VI) concentrations. The strain 8 tolerated a 6% (w/v) NaCl concentration was able to bioaccumulate more than 95% of the applied chromium(VI) at the 97.6–224.4 mg l⁻¹ initial chromium(VI) concentrations. The results presented in this paper was shown that these pure and mixed cultures might be of use for the bioaccumulation of chromium(VI) from saline wastewater.     

Effect of sulfate and iron on physico-chemical characteristics of anaerobic granular sludge

This research investigated the effect of the substrate composition (no substrate, glucose, glucose + sulfate or glucose + sulfate + iron) on the physico-chemical characteristics of two different anaerobic granular sludges as a function of time. The sludges were fed batch wise (pH 7, 30 °C) at an organic loading rate of 1.2 g COD l⁻¹ d⁻¹ (0.04 g COD g VSS⁻¹ day⁻¹) for 30 days. The presence of sulfate (COD/sulfate ratio = 1) in the feed of glucose fed anaerobic sludges did not change the physico-chemical characteristics throughout the incubation. In contrast, the presence of iron in the feed (in addition to glucose and sulfate, COD/iron ratio = 1) reduced the protein and carbohydrate content in the SMP and EPS with about 50% after 30 days incubation compared to the other feeding conditions. The sludge grown on glucose + sulfate + iron contained much more iron (+300–500%) and sulfur (+200–350%) than the other incubated sludges both after 14 and 30 days. The higher mineral content (lower VSS content) and the decrease of the EPS content contributed to the disintegration of iron fed granules, as shown by their lower size particles. However, the iron fed sludge displayed a higher granule strength than the other incubated sludges. Although an appreciable variation in the granule strength was noticed between the sludges investigated, it was not possible to relate these differences to their inorganic composition, the chemical composition of the extracted polymers or to the physical characteristics investigated.     

Pilot-scale production of 1,3-propanediol using Klebsiella pneumoniae

The conversion of glycerol to 1,3-propanediol (PDO) using Klebsiella pneumoniae M5al under anaerobic condition was scaled up from scale 5 to 5000 l in series. A simple strategy for scale-up was to transfer the optimized conditions of a lab scale bioreactor to pilot-scale fermentation. Multistage inocula were developed and their fermentation abilities were assessed in a small-scale fermenter. The experimental results showed that inoculum development in the early steps of a scale-up process could influence the outcomes of a large scale fermentation. Through three-stage liquid inoculum development and a pulse addition of (NH₄)₂SO₄ and yeast extract at 30 h of fermentation, the best results in a 5000 l fermentation were achieved leading to 58.8 g l⁻¹ 1,3-propanediol with a yield of 0.53 mol mol⁻¹ glycerol and productivity of 0.92 g l⁻¹ h⁻¹. This is the first report on pilot-scale 1,3-propanediol production using K. pneumoniae.     

Extent estimates and land cover relationships for functional indicators in non-wadeable rivers

Functional indicators are being increasingly used to assess waterway health but their responses to pressure in non-wadeable rivers have not been widely documented or applied in modern survey designs that provide unbiased estimates of extent. This study tests the response of river metabolism and loss in cotton strip tensile strength across a land use pressure gradient in non-wadeable rivers of northern New Zealand, and reports extent estimates for river metabolism and decomposition rates. Following adjustment for probability of selection, ecosystem respiration (ER) and gross primary production (GPP) for the target population of order 5–7 non-wadeable rivers averaged −7.3 and 4.8 g O₂ m⁻² d⁻¹, respectively, with average P/R < 1 indicating dominance by heterotrophic processes. Ecosystem respiration was <−3.3 g O₂ m⁻² d⁻¹ for 75% of non-wadeable river length with around 20% of length between −10 and −20 g O₂ m⁻² d⁻¹. Cumulative distribution functions of cotton strength loss estimates indicated a more-or-less linear relationship with river km reflecting an even spread of decay rates (range in k 0.0007–0.2875 d⁻¹) across non-wadeable rivers regionally. A non-linear relationship with land cover was detected for GPP which was typically <5 g O₂ m⁻² d⁻¹ where natural vegetation cover was below 20% and greater than 80% of upstream catchment area. For cotton strength loss, the relationship with land cover was wedge-shaped such that sites with >60% natural cover had low decay rates (<0.02 d⁻¹) with variability below this increasing as natural cover declined. Using published criteria for assessing waterway health based on ER and GPP, 232–298 km (20–29%) of non-wadeable river length was considered to have severely impaired ecosystem functioning, and 436–530 km (42–50%) had no evidence of impact on river metabolism.     

Operation of a bioelectrochemical system as a polishing stage for the effluent from a two-stage biohydrogen and biomethane production process

Anaerobic bioenergy production processes including fermentative biohydrogen (BioH₂), anaerobic digestion (AD) and bioelectrochemical system have been investigated for converting municipal waste or various biomass feedstock to useful energy carriers. However, the performance of a microbial fuel cell (MFC) fed on the effluent from a two-stage biogas production process has not yet been investigated extensively in continuous reactor operation on complex substrates. In this study we have investigated the extent to which a microbial fuel cell (MFC) can reduce COD and recover further energy from the effluent of a two-stage biohydrogen and biomethane system. The performance of a four-module tubular MFC was determined at six different organic loadings (0.036–6.149 g sCOD L⁻¹ d⁻¹) in terms of power generation, COD removal efficiency, coulombic efficiency (CE) and energy conversion efficiency (ECE). A power density of 3.1 W m⁻³ was observed at the OLR = 0.572 g sCOD L⁻¹ d⁻¹, which resulted in the highest CE (60%) and ECE (0.8%), but the COD removal efficiency decreased at higher organic loading rates (35.1–4.4%). The energy recovery was 92.95 J L⁻¹ and the energy conversion efficiency, based on total influent COD was found to be 0.48–0.81% at 0.572 g sCOD L⁻¹ d⁻¹. However, the energy recovery by the MFC is only reported for a four-module reactor and improved performance can be expected with an extended module count, as chemical energy remained available for further electrogenesis.     

Multiple regeneration pathways in Spathoglottis plicata Blume — A study in vitro

Asymbiotic germination of immature seeds (embryos), and mature seeds and micropropagation of Spathoglottis plicata were described. Effects of three nutrition media namely, Murashige & Skoog (MS); Phytamax (PM); and Phyto-Technology orchid seed sowing medium (P₇₂₃), two carbon sources such as glucose and sucrose at 2–3% (w/v), two plant growth regulators such as 6-benzylaminopurine (BAP; 0.5–3.0 mg l^− 1) and α-naphthalene acetic acid (NAA; 0.5–2.0 mg l^− 1) and peptone (2.0 g l^− 1) were examined on seed germination, early protocorm development and micropropagation. The maximum germination of mature seeds (95%) was recorded in PM medium supplemented with 2% (w/v) sucrose + 2.0 g l^− 1 peptone. For germination of embryos P₇₂₃ medium supplemented with 1.0 mg l^− 1 BAP proved best. Multiple shoot buds or protocorm-like bodies (PLBs) were produced from stem segments of in vitro raised seedlings. Both direct organogenesis and embryogenesis were observed and the morphogenetic response was initiated by different concentrations and combinations of PGRs. The optimum PGR combination for maximal PLB regeneration was 1.0 mg l^− 1 NAA + 2.5 mg l^− 1 BAP, while 1.0 mg l^− 1 NAA + 1.0 mg l^− 1 BAP for shoot bud development. Strong and stout root system was induced in half strength PM medium supplemented with 0.5 mg l^− 1 IAA. The well-rooted plantlets were transferred to pots containing a potting mixture composed of saw dust, coconut coir, humus, and coal pieces at 1:1:1:2 (w/w) with 80% survival in outside environment and flowered after two years of transfer.     

Enhancement of polysaccharide production in suspension cultures of protocorm-like bodies from Dendrobium huoshanense by optimization of medium compositions and feeding of sucrose

A strategy that optimization of medium compositions for maximum biomass followed by feeding of sucrose for maximum polysaccharide synthesis was developed for enhancing polysaccharide production in suspension culture of protocorm-like bodies (PLBs) of Dendrobium huoshanense C.Z. Tang et S.J. Cheng. In growth stage, the original half-strength MS medium was optimized with carbon sources, nitrogen sources and metal ion combinations. The effects of different carbon sources on PLBs growth were remarkable and sucrose at 35 g l⁻¹ was the most suitable. Sole nitrate nitrogen of 30 mmol l⁻¹ was the best for PLBs growth. Metal ions (Ca²⁺, Fe²⁺, Mn²⁺ and Zn²⁺) showed different influences on PLBs growth. The optimal concentration of Ca²⁺, Fe²⁺, Mn²⁺ and Zn²⁺ was 4.5 mmol l⁻¹, 0.1 mmol l⁻¹, 0.5 mmol l⁻¹ and 0.06 mmol l⁻¹, respectively. In the optimized medium (sucrose, nitrate, Ca²⁺, Fe²⁺, Mn²⁺ and Zn²⁺ concentration as described above, the other component concentration seen in half-strength MS), 33.9 g DW l⁻¹ PLBs were harvested after 30 days of culture and biomass increase was improved 245% as compared with that in the original medium. In production stage, polysaccharide synthesis was significantly improved by the feeding sucrose. The maximum polysaccharide production (22 g l⁻¹) was obtained in the case of 50 g l⁻¹ sucrose feeding at day 30 of culture, which was about 109-fold higher than that in the original medium without feeding of sucrose.     

Evaluation of distant carbon sources in biosurfactant production by a gamma ray-induced Pseudomonas putida mutant

Pseudomonas putida 33 wild strain, subjected to gamma ray mutagenesis and designated as P. putida 300-B mutant was used as microbial rhamnolipid-producer by using distant carbon sources (viz. hydrocarbons, waste frying oils ‘WFOs’, vegetable oil refinery wastes and molasses) in the minimal media under shake flask conditions. The behavior of glucose as co-substrate and growth initiator was examined. The 300-B mutant strain showed its ability to grow on all the substrates tested and produced rhamnolipid surfactants to different extents however; soybean and corn WFOs were observed to be preferred carbon sources followed by kerosene and paraffin oils, respectively. The best cell biomass (3.5 g l⁻¹) and rhamnolipids yield (4.1 g l⁻¹) were obtained with soybean WFO as carbon source and glucose as growth initiator under fed-batch cultivation showing an optimum specific growth rate (μ) of 0.272 h⁻¹, specific product yield (q_p) of 0.318 g g⁻¹ h and volumetric productivity (P_V) of 0.024 g l⁻¹ h. The critical micelle concentration of its culture supernatant was observed to be 91 mg rhamnolipids l⁻¹ and surface tension as 31.2 mN m⁻¹.     

Investigation on the properties and kinetics of glucose-fed aerobic granular sludge

Aerobic granulation is a process in which suspended biomass aggregate and form discrete well-defined granules in aerobic systems. To investigate the properties and kinetics of aerobic granular sludge, aerobic granules were cultivated with glucose synthetic wastewater in a series of sequencing batch reactors (SBR). The spherical shaped granules were observed on 8th day with the mean diameter of 0.1 mm. With the organic loading rate (OLR) being increased to 4.0 g COD L⁻¹ d⁻¹, aerobic granules grew matured with spherical shape. The size of granules ranged from 1.2 to 1.8 mm, and the corresponding settling velocity of individual granule was 24.2–36.4 m h⁻¹. The oxygen utilization rate (OUR) of mature granules was 41.90 g O₂ kg MLSS⁻¹ h⁻¹, which was two times higher than that of activated sludge (18.32 g O₂ kg MLSS⁻¹ h⁻¹). The experimental data indicated that the substrate utilization and biomass growth kinetics generally followed Monod's kinetics model. The corresponding kinetic coefficients of k (maximum specific substrate utilization rate), K_s (half velocity coefficient), Y (growth yield coefficient) and K_d (decay coefficient) were determined as follows, k_c = 23.65 d⁻¹, K_c = 3367.05 mg L⁻¹, K_N = 0.038 d⁻¹, K_N = 29.65 mg L⁻¹, Y = 0.1927–0.2022 mg MMLS (mg COD)⁻¹ and K_d = 0.00845–0.0135 d⁻¹, respectively. Those properties of aerobic granules made aerobic granules system had a short setup period, high substrate utilization rate and low sludge production.     

Uptake and allocation of 13C by Enhalus acoroides at sites differing in light availability

Carbon fixation and allocation were studied using ¹³C incubation and leaf marking techniques in mature monospecific stands of Enhalus acoroides L.f. Royle in August 1998 and January 1999 in Banten Bay, Indonesia. The highest rate of ¹³C uptake (>0.008 g ¹³C g C⁻¹ d⁻¹) was found in the middle to distal parts of leaves of E. acoroides. Young and senescing leaves numbers had lower ¹³C incorporation compared to mature leaves. The incorporation of ¹³C by epiphytes on the leaves was higher than that of the leaves themselves (>0.01 g ¹³C g C⁻¹ d⁻¹). The results showed that turbidity of the water influenced the leaf growth, productivity and Relative Growth Rate of E. acoroides, which were lower at Kepuh Island, the more turbid site. However, at Kepuh Island, where the water column was turbid, the plant could still harvest sufficient light for an uptake rate of ¹³C, higher than the uptake rates at Kubur and Panjang Islands, stations with a much more transparent water column (on average 0.0047 g ¹³C g C⁻¹ d⁻¹ at Kepuh Island, versus 0.0045 g ¹³C g C⁻¹ d⁻¹ at Panjang Island and 0.0034 g ¹³C g C⁻¹ d⁻¹ at Kubur Island). There was evidence that ¹³C was exported from the incubated shoots to the roots and rhizomes and to neighboring shoots of E. acoroides in clear water, but not in turbid water. We suggest that single shoots of E. acoroides are able to grow in turbid water under low light conditions. They assimilate sufficient carbon for their own maintenance but are not able to export to neighboring plant parts.     

Over-production of human interferon-γ by HCDC of recombinant Escherichia coli

A simple fed-batch process was developed using a modified variable specific growth rate feeding strategy for high cell density cultivation of Escherichia coli BL21 (DE3) expressing human interferon-gamma (hIFN-γ). The feeding rate was adjusted to achieve the maximum attainable specific growth rate during fed-batch cultivation. In this method, specific growth rate was changed from a maximum value of 0.55 h⁻¹ at the beginning of feeding and then it was reduced to 0.4 h⁻¹ at induction time.The final concentration of biomass and IFN-γ was reached to ∼115 g l⁻¹ (DCW) and 42.5 g(hIFN-γ) l⁻¹ after 16.5 h, also the final specific yield and overall productivity of recombinant hIFN-γ (rhIFN-γ) were obtained 0.37 g(hIFN-γ) g⁻¹ DCW and 2.57 g(hIFN-γ) l⁻¹ h⁻¹, respectively. According to available data this is the highest specific yield and productivity that has been reported for recombinant proteins production yet.     

Poly(3-hydroxybutyrate) production by Cupriavidus necator using waste glycerol

Polyhydroxyalkanoates (PHAs) have been recognized as good substitutes for the non-biodegradable petrochemically produced polymers. However, their high (real or estimated) current production cost limits their industrial applications. This work exploits two strategies to enhance PHAs substitution potential: the increase in PHA volumetric productivity in high density cultures and the use of waste glycerol (GRP), a by-product from the biodiesel industry, as primary carbon source for cell growth and polymer synthesis. Cupriavidus necator DSM 545 was used to accumulate poly(3-hydroxybutyrate) (P(3HB)) from GRP and from commercial glycerol (PG) as control substrate. On PG, productivities between 0.6 g_PHB L^?1 h^?1 and 1.5 g_PHB L^?1 h^?1 were attained. The maximum cell DW was 82.5 g_DW L^?1, the P(3HB) content being 62%. When GRP was used, 68.8 g_DW L^?1 with a P(3HB) accumulation of 38% resulting in a final productivity of 0.84 g_PHB L^?1 h^?1 was obtained. By decreasing the biomass concentration at which accumulation was triggered, a productivity of 1.1 g_PHB L^?1 h^?1 (50% P(3HB), w/w) was attained using GRP. P(3HB) molecular weights (M_w) ranged from 7.9 × 10⁵ to 9.6 × 10⁵ Da.     

Treatment of dye-rich wastewater by an immobilized thermophilic cyanobacterial strain: Phormidium sp.

The removal of Remazol Blue and Reactive Black B by the immobilized thermophilic cyanobacterial strain Phormidium sp. was investigated under thermophilic conditions in a batch system, in order to determine the optimal conditions required for the highest dye removal. In the experiments, performed at pH 8.5, with different initial dye concentrations between 9.1 mg l⁻¹ and 82.1 mg l⁻¹ and at 45 °C, calcium alginate immobilized Phormidium sp. showed high dye decolorization, with maximum uptake yields ranging from 50% to 88% at all dye concentrations tested. When the effects of high dye concentrations on dye removal were investigated, the highest uptake yield in the beads was 50.3% for 82.1 mg l⁻¹ Remazol Blue and 60.0% for 79.5 mg l⁻¹ Reactive Black B. The highest color removal was detected at 45 °C and 50 °C incubation temperatures for all dye concentrations. As the temperature decreased, the removal yield of immobilized Phormidium sp. also decreased. At about 75 mg l⁻¹ initial dye concentrations, the highest specific dye uptake measured was 41.29–41.17 mg g⁻¹ for Remazol Blue and 47.69–43.82 mg g⁻¹ for Reactive Black B at 45 °C and 50 °C incubation temperatures, respectively, after 8 days incubation.     

Enzymatic hydrolysis of corncob and ethanol production from cellulosic hydrolysate

 Enzymatic hydrolysis of corncob and ethanol fermentation from cellulosic hydrolysate were investigated. After corncob was pretreated by 1% H₂SO₄ at 108 °C for 3 h, the cellulosic residue was hydrolyzed by cellulase from Trichoderma reesei ZU-02 and the hydrolysis yield was 67.5%. Poor cellobiase activity in T. reesei cellulase restricted the conversion of cellobiose to glucose, and the accumulation of cellobiose caused severe feedback inhibition to the activities of β-1,4-endoglucanase and β-1,4-exoglucanase in cellulase system. Supplementing cellobiase from Aspergillus niger ZU-07 greatly reduced the inhibitory effect caused by cellobiose, and the hydrolysis yield was improved to 83.9% with enhanced cellobiase activity of 6.5 CBU g⁻¹ substrate. Fed-batch hydrolysis process was started with a batch hydrolysis containing 100 g l⁻¹ substrate, with cellulosic residue added at 6 and 12 h twice to get a final substrate concentration of 200 g l⁻¹. After 60 h of reaction, the reducing sugar concentration reached 116.3 g l⁻¹ with a hydrolysis yield of 79.5%. Further fermentation of cellulosic hydrolysate containing 95.3 g l⁻¹ glucose was performed using Saccharomyces cerevisiae 316, and 45.7 g l⁻¹ ethanol was obtained within 18 h. The research results are meaningful in fuel ethanol production from agricultural residue instead of grain starch.     

Protocorm culture of Dendrobium candidum in balloon type bubble bioreactors

Protocorm cultures of Dendrobium candidum were established in balloon type bubble bioreactors using Murashige and Skoog (MS) medium with 0.5 mg l⁻¹ α-naphthaleneacetic acid (NAA), 2.5% (w/v) sucrose, 5:25 mM NH₄:NO₃ and 1% (v/v) banana homogenate for the production of biomass and bioactive compounds. In 3 l bioreactor containing 2 l medium, a maximum protocorm biomass (21.0 g l⁻¹ dry biomass) and also optimum quantities of total polysaccharides (389.3 mg g⁻¹ DW), coumarins (18.0 mg g⁻¹ DW), polyphenolics (11.9 mg g⁻¹ DW), and flavonoids (4.5 mg g⁻¹ DW) were achieved after 7 weeks of culture. Based on these studies, 5 and 10 l bioreactor cultures were established to harvest 80 g and 160 g dry biomass. In 10 l bioreactors, the protocorms grown were accumulated with optimal levels of polysaccharides (424.1 mg g⁻¹ DW), coumarins (15.8 mg g⁻¹ DW), polyphenols (9.03 mg g⁻¹ DW) and flavonoids (4.7 mg g⁻¹ DW). The bioreactor technology developed here will be useful for the production of important bioactive compounds from D. candidum.     

Characterization of a recombinant Escherichia coli TOP10 [pQR239] whole-cell biocatalyst for stereoselective Baeyer–Villiger oxidations

This paper describes the kinetic characterization of a recombinant whole-cell biocatalyst for the stereoselective Baeyer–Villiger type oxidation of bicyclo[3.2.0]hept-2-en-6-one to its corresponding regio-isomeric lactones (−)-(1S,5R)-2-oxabicyclo[3.3.0]oct-6-en-3-one and (−)-(1R,5S)-3-oxabicyclo[3.3.0]oct-6-en-2-one. Escherichia coli TOP10 [pQR239], expressing cyclohexanone monooxygenase (CHMO) from Acinetobacter calcoaceticus (NCIMB 9871), was shown to be suitable for this biotransformation since it expressed CHMO at a high level, was simple to produce, contained no contaminating lactone hydrolase activity and allowed the intracellular recycle of NAD(P)H necessary for the biotransformation. A small-scale biotransformation reactor (20 ml) was developed to allow rapid collection of intrinsic kinetic data. In this system, the optimized whole-cell biocatalyst exhibited a significantly lower specific lactone production activity (55–60 μmol min⁻¹ g⁻¹ dry weight) than that of sonicated cells (500 μmol min⁻¹ g⁻¹ dry weight). It was shown that this shortfall was comprised of a difference in the pH optima of the two biocatalyst forms and mass transfer limitations of the reactant and/or product across the cell barrier. Both reactant and product inhibition were evident. The optimum ketone concentration was between 0.2 and 0.4 g l⁻¹ and at product concentrations above 4.5–5 g l⁻¹ the specific activity of the whole cells was zero. These results suggest that a reactant feeding strategy and in situ product removal should be considered in subsequent process design.