Repeated Batch Culture Research Articles

Utilization of agro-industrial byproducts as low-cost nutrient sources for production of kefiran and lactic acid by Lactobacillus kefiranofaciens

Kefiran is an exopolysaccharide derived from kefir grains that has antioxidant and antimicrobial properties. The high production cost of kefiran is primarily attributed to the expensive fermentation media. Therefore, the use of byproducts from agro-industries could present an economically favorable alternative. In this study, locally available mature coconut water (MCW) and whey lactose (WL) were used as cost-effective nutrient sources for kefiran production by Lactobacillus kefiranofaciens JCM 6985. MCW gave a higher yield of kefiran and acid than WL and was comparable to the commercial medium. The optimal sugar concentration for kefiran and acid production from MCW was 30 g/L, and the pH control during cultivation could alleviate the inhibitory effect of acidic pH and enhance kefiran and acid production. When the MCW was added with yeast extract at 3 g-nitrogen/L, the kefiran and acid production increased up to 3.26 ± 0.03 g/L and 15.5 ± 0.7 g/L, respectively. Moreover, the kefiran and acid production was scaled up in the bioreactor and enhanced by the repeated-batch cultivation for six cycles. The structure analysis indicated that kefiran produced by L. kefiranofaciens when using MCW had a similar structure to that extracted from kefir grains. The findings demonstrate promising methods for cost-effective production of kefiran and lactic acid using inexpensive nutrient sources such as MCW.

Efficient conversion of cane molasses into Tremella fuciformis polysaccharides with enhanced bioactivity through repeated batch culture

Tremella fuciformis polysaccharide (TFPS) is a natural mushroom mucopolysaccharide widely used in health foods, medical care, cosmetic and surgical materials. In this study, we developed an efficient strategy for the repeated batch production of highly bioactive TFPS from the agro-industrial residue cane molasses. Cane molasses contained 39.92 % sucrose (w/w), 6.36 % fructose and 3.53 % glucose, all of which could be utilized by T. fuciformis spores, whereas, the TFPS production efficiency only reached 0.74 g/L/d. Corn cobs proved to be the best immobilized carrier that could tightly absorb spores and significantly shorten the fermentation lag period. The average yield of TFPS in eight repeated batch culture was 5.52 g/L with a production efficiency of 2.04 g/L/d. The average fermentation cycle after optimization was reduced by 61.61 % compared with the initial conditions. Compared to glucose as a carbon source, cane molasses significantly increased the proportion of low-molecular-weight TFPS (TFPS-2) in total polysaccharides from 3.54 % to 17.25 % (w/w). Moreover, TFPS-2 exhibited potent antioxidant capacity against four free radicals (O2−, ABTS+, OH, and DPPH). In conclusion, this study lays the foundation for the efficient conversion of cane molasses and production of TFPS with high bioactivity.

Magnetite-boosted syntrophic conversion of acetate to methane during thermophilic anaerobic digestion.

Using a batch thermophilic anaerobic system established with 60 mL serum bottles, the mechanism on how microbial enrichments obtained from magnetite-amended paddy soil via repeated batch cultivation affected methane production from acetate was investigated. Magnetite-amended enrichments (MAEs) can improve the methane production rate rather than the methane yield. Compared with magnetite-unamended enrichments, the methane production rate in MAE was improved by 50%, concomitant with the pronounced electrochemical response, high electron transfer capacity, and fast acetate degradation. The promoting effects might be ascribed to direct interspecies electron transfer facilitated by magnetite, where magnetite might function as electron conduits to link the acetate oxidizers (Anaerolineaceae and Peptococcaceae) with methanogens (Methanosarcinaceae). The findings demonstrated the potential application of MAE for boosting methanogenic performance during thermophilic anaerobic digestion.

Extended Biosynthesis of Rhamnolipid by Immobilized Pseudomonas aeruginosa USM-AR2 Cells in a Fluidized Bed Bioreactor.

This study aims to evaluate rhamnolipid production by immobilized Pseudomonas aeruginosa USM-AR2 cells using waste cooking oil (WCO) as the carbon source. P. aeruginosa USM-AR2 cells were entrapped in polyvinyl alcohol (PVA)-alginate hydrogel beads. The performance of entrapped cells was compared with free cells in shake flasks before cultivation in a custom-designed fluidized bed reactor (FBR). A mass of 1 g wet cells entrapped in PVA-alginate hydrogel beads were successfully recycled 3 times in shake flasks at 200 rpm, producing between 0.66-1.34 g L-1 rhamnolipid after 120 h. Meanwhile, cultivation of entrapped cells in FBR with broth recirculation showed that the suitable hydrogel beads to medium ratio was 1:20 ratio at an aeration rate of 0.5 vvm, producing between 0.77 to 1.58 g L-1 rhamnolipid, degrading 8.67 to 20.93 g L-1 of waste cooking oil in 15 cycles of repeated batch cultivation. Entrapped P. aeruginosa USM-AR2 cells achieved a higher rhamnolipid production, by 1.03-fold during cycle 3 in shake flasks and 1.19-fold during cycle 11 in an FBR, compared to free cells. These results show that entrapment enables reusability of viable cells and maintains stability of rhamnolipid production throughout the extended cultivation, increasing cell tolerance to perturbations in fermentation conditions.

Biotechnological enhancement of lactic acid conversion from pretreated palm kernel cake hydrolysate by Actinobacillus succinogenes 130Z

The aim of this study was to establish an improved pretreatment and fermentation method i.e. immobilized cells for high recovery of fermentable sugars from palm kernel cake (PKC) and its effects on fermentability performance by Actinobacillus succinogenes 130Z in the conversion of the fermentable sugar to lactic acid. The effects of oxalic acid concentrations (1–6% w/v) and residence times (1–5 h) on the sugar recovery were initially investigated and it was found that the highest mannose concentration was 25.1 g/L at the optimum hydrolysis conditions of 4 h and 3% (w/v) oxalic acid. The subsequent enzymatic saccharification of the pretreated PKC afforded the highest enzymatic digestibility with the recovered sugars amounting to 25.18 g/L and 9.14 g/L of mannose and glucose, respectively. Subsequently, the fermentability performance of PKC hydrolysate was evaluated and compared in terms of cultivation phases (i.e. mono and dual-phases), carbonate loadings (i.e. magnesium and sodium carbonates), and types of sugars (i.e. glucose and mannose). The highest titer of 19.4 g/L lactic acid was obtained from the fermentation involving A. succinogenes 130Z in dual-phase cultivation supplemented with 30 g/L of magnesium carbonate. Lactic acid production was further enhanced by using immobilized cells with coconut shell-activated carbon (CSAC) of different sizes (A, B, C, and D) in the repeated batch cultivation of dual-phase fermentation producing 31.64 g/L of lactic acid. This work sheds light on the possibilities to enhance the utilization of PKC for lactic acid production via immobilized A. succinogenes 130Z.

Tapping on the Potential of Hyaluronic Acid: from Production to Application.

The manufacture, purification, and applications of hyaluronic acid (HA) are discussed in this article. Concerning the growing need for affordable, high-quality HA, it is essential to consider diverse production techniques using renewable resources that pose little risk of cross-contamination. Many microorganisms can now be used to produce HA without limiting the availability of raw materials and in an environmentally friendly manner. The production of HA has been associated with Streptococci A and C, explicitly S. zooepidemicus and S. equi. Different fermentation techniques, including the continuous, batch, fed-batch, and repeated batch culture, have been explored to increase the formation of HA, particularly from S. zooepidemicus. The topic of current interest also involves a complex broth rich in metabolites and residual substrates, intensifying downstream processes to achieve high recovery rates and purity. Although there are already established methods for commercial HA production, the anticipated growth in trade and the diversification of application opportunities necessitate the development of new procedures to produce HA with escalated productivity, specified molecular weights, and purity. In this report, we have enacted the advancement of HA technical research by analyzing bacterial biomanufacturing elements, upstream and downstream methodologies, and commercial-scale HA scenarios.

Producción de etanol a partir de desechos frutales mexicanos utilizando una nueva cepa de Saccharomyces cerevisiae

In the Mexican croplands are generated large amounts of agroindustrial wastes that are usually not exploited. Damaged fruits wasted in the municipality of Tres Valles, Veracruz, are an excellent feedstock to produce ethanol, since do not need a sophisticated pretreatment and have high fermentable sugar concentrations. In this work is described ethanol production from damaged fruits by a new strain of Saccharomyces cerevisiae isolated from agave sp. wastes. Fermentations were carried out in batch and repeated batch cultures using biocatalysts formed by S. cerevisiae AP1 cells immobilized into alginate-coated polyester fiberfill. Biocatalysts showed a high fermentative capability at reducing sugar concentrations higher than 30 g L-1.  In batch cultures, with 32.58 g reducing sugar L-1, was produced up to 15.39 g ethanol L-1 at 16 h, with a volumetric productivity of 0.962 g L-1 h-1 and a fermentation efficiency of 94.77%. Instead in a 5-cycle repeated batch fermentation, with a reducing sugar content among 30 to 43 g L-1, ethanol production in each cycle was fast, higher than 15 g L-1, with fermentation efficiencies higher than 80%, and with volumetric productivities from 2.5 to 2.9 g L-1 h-1 after second cycle. Afterwards five cycles of repeated batch fermentation, total ethanol production was 95.41 g L-1 in just 44 h process.

Effects of recycling hyper-thermal inoculum by repeated batch cultivation into co-composting of sludge and livestock-poultry manure

The enrichment and adaptation of hyper-thermal compost-derived thermophilic inoculum by repeated batch cultivation (RBC) was conducted by investigating bacterial community. The effects of recycling hyper-thermal inoculum by RBC into co-composting were investigated through evaluating the influences of temperature, pH, moisture, C/N ratio, transformation of nitrogen, composting maturity, humification levels and scanning electron microscopy (SEM). The results showed that RBC enriched the thermophilic bacterial community and nitrogen fixation bacteria of the compost-derived thermophilic inoculum. Simultaneously, recycling the inoculum into co-composting increased the temperature, nitrate nitrogen (NO3 –-N) and Germination index (GI), and improved the transformation of nitrogen and humification levels. Conclusively, recycling hyper-thermal inoculum by RBC into co-composting can improve the degradation process.

Comparative study of bioethanol production from bagasse pith by Pichia stipitis in free cell system and immobilized electrosprayed micro-beads: SSF and combined hydrolysates fermentation

Biofuels production from biomass are receiving special attention due to the availability of raw materials, suitable conversion yield and renewability. The primary objective of this study was comparison of bioethanol productivity from bagasse pith by free and immobilized Pichia stipitis. Alginate micro-beads at various sizes fabricated by electrospray technique. At the first stage, bioethanol production using free cells in two processes including fermentation of combined acid and enzymatic hydrolysates and simultaneous saccharification and fermentation (SSF) was optimized. In fermentation of combined hydrolysates, maximum production equal to 9.53 and 12.68 g/L was observed by free and immobilized cells, respectively at cell concentration 6 %, portion of acid hydrolysate 40 % and total sugar concentration 50 %. Besides, maximum production in SSF equal to 9.93 and 10.62 g/L was obtained by free and immobilized cells, respectively at 6 % cell, 4 g solid and enzyme content of 50 FPU/g solid. However, in SSF fermentation, the beads size had no significant effect on ethanol production while in combined hydrolysates fermentation, bioethanol production considerably affected by the beads size and 13.9 g/L ethanol was obtained in minimum beads size. Additionally, long-term stability of beads was examined in repeated-batch cultures, showing effective bioethanol production up to five reuse cycles.

Effectiveness of tetrahydrofuran at enhancing the 1,4-dioxane degradation ability of activated sludge lacking prior exposure to 1,4-dioxane.

1,4-dioxane (DX) is a contaminant of emerging concern in water environments. The enrichment of DX-degrading bacteria indigenous to activated sludge is key for the efficient biological removal of DX in wastewater. To identify an effective substrate, which enables the selective enrichment of DX-degrading bacteria and has lower toxicity and persistence than DX, this study explored the effectiveness of tetrahydrofuran (THF) at enhancing the DX degradation ability of activated sludge without historical exposure to DX. Although the activated sludge initially exhibited negligible ability to degrade DX (100 mg-C/L) as the sole carbon source, the repeated batch cultivation on THF could enrich bacterial populations capable of degrading DX, inducing the DX degradation ability in activated sludge as effectively as DX did. The THF-enrichment culture after 4 weeks degraded 100 mg-C/L DX almost completely within 21 d. Sequencing analyses revealed that soluble di-iron monooxygenase group 5C, including THF/DX monooxygenase, would play a dominant role in the initial oxidation of DX in THF-enrichment culture, which completely differed from the enrichment culture cultivated on DX. The results indicate that THF can be applied as an effective substrate to enhance the DX degradation ability of microbial consortia, irrespective of the intrinsic ability.

Production of Carotenoids and Phospholipids by Thraustochytrium sp. in Batch and Repeated-Batch Culture.

The carotenogenic thraustochytrid Thraustochytrium sp. RT2316-16 was grown in batch and repeated-batch cultures using different feeds containing glucose, or glycerol, and yeast extract, for the production of lipids, phospholipids and carotenoids. RT2316-16 produced canthaxanthin, astaxanthin and β-carotene. The effects of biotin, ascorbic acid, light and temperature were evaluated in some of the experiments. In 2-day-old batch cultures, the combined mass percentage of eicosapentaenoic acid and docosahexaenoic acid in total lipids was between 16.5% (glycerol-based medium in the dark; biomass concentration = 4.2 ± 1.1 g L−1) and 42.6% (glucose-based medium under light; biomass concentration = 3.3 ± 0.1 g L−1), decreasing to 3.8% and 6.1%, respectively, after day 4. In repeated-batch cultures, the total lipids in the biomass increased after glucose or glycerol was fed alone, whereas the total carotenoids (168 ± 7 μg g−1 dry weight (DW)) and phospholipids in the biomass increased after feeding with yeast extract. The biomass with the highest content of phospholipids (28.7 ± 4.3 mg g−1 DW) was obtained using a feed medium formulated with glycerol, yeast extract and ascorbic acid. Glycerol was the best carbon source for the production of a biomass enriched with total lipids (467 ± 45 mg g−1 DW). The composition of carotenoids depended strongly on the composition of the feed. Repeated-batch cultures fed with yeast extract contained canthaxanthin as the main carotenoid, whereas in the cultures fed only with glucose, the biomass contained mainly β-carotene.

Enhanced l-Malic Acid Production by Aspergillus oryzae DSM 1863 Using Repeated-Batch Cultivation.

l-Malic acid is a C4-dicarboxylic acid and a potential key building block for a bio-based economy. At present, malic acid is synthesized petrochemically and its major market is the food and beverages industry. In future, malic acid might also serve as a building block for biopolymers or even replace the commodity chemical maleic anhydride. For a sustainable production of l-malic acid from renewable resources, the microbial synthesis by the mold Aspergillus oryzae is one possible route. As CO2 fixation is involved in the biosynthesis, high yields are possible, and at the same time greenhouse gases can be reduced. In order to enhance the production potential of the wild-type strain Aspergillus oryzae DSM 1863, process characteristics were studied in shake flasks, comparing batch, fed-batch, and repeated-batch cultivations. In the batch process, a prolonged cultivation time led to malic acid consumption. Keeping carbon source concentration on a high level by pulsed feeding could prolong cell viability and cultivation time, however, did not result in significant higher product levels. In contrast, continuous malic acid production could be achieved over six exchange cycles and a total fermentation time of 19 days in repeated-batch cultivations. Up to 178 g/L l-malic acid was produced. The maximum productivity (0.90 ± 0.05 g/L/h) achieved in the repeated-batch cultivation had more than doubled than that achieved in the batch process and also the average productivity (0.42 ± 0.03 g/L/h for five exchange cycles and 16 days) was increased considerably. Further repeated-batch experiments confirmed a positive effect of regular calcium carbonate additions on pH stability and malic acid synthesis. Besides calcium carbonate, nitrogen supplementation proved to be essential for the prolonged malic acid production in repeated-batch. As prolonged malic acid production was only observed in cultivations with product removal, product inhibition seems to be the major limiting factor for malic acid production by the wild-type strain. This study provides a systematic comparison of different process strategies under consideration of major influencing factors and thereby delivers important insights into natural l-malic acid production.

Isopropanol-butanol-ethanol production by cell-immobilized vacuum fermentation

The Isopropanol-Butanol-Ethanol productivity by solventogenic clostridia can increase when cells are immobilized on low-cost, renewable fibrous materials; however, butanol inhibition imposes the need for dilute sugar solutions (less than40 g/L). To alleviate this problem, the in-situ vacuum product recovery technique was applied to recover IBE in repeated-batch cultivation of Clostridium beijerinckii DSM 6423 immobilized on sugarcane bagasse. Five repeated batch cycles were conducted in a 7-L bioreactor containing P2 medium (∼60 g/L glucose) and bagasse packed in 3D-printed concentric annular baskets. In three cycles, glucose was consumed by 86% on average, the IBE productivity was 0.35 g/L∙h or 30% and 17% higher relative to free- and immobilized (without vacuum)-cell cultures. Notably, the product stream contained 45 g/L IBE. However, the fermentation was unsatisfactory in two cycles. Finally, by inserting a fibrous bed with hollow annuli in a vacuum fermentation, this work introduces the concept of an internal-loop boiling-driven fibrous-bed bioreactor.

Autotrophic and mixotrophic biomass production of the acidophilic Galdieria sulphuraria ACUF 64

Abstract Galdieria sulphuraria is an acidophilic microalga isolated in proximity of sulfuric ponds where pH is below 3 and most organisms cannot grow. We cultivated G. sulphuraria ACUF 64 free of contamination for over 2 months in a medium containing organic carbon at pH 1.7 with continuous, high intensity, lighting. We compared biomass productivity of chemostat and repeated batch cultivations. The optimal biomass density in autotrophic and mixotrophic cultures was identified. In autotrophy biomass productivity was 28.3 g x ·m −2 ·day −1 , 1.8 to 7.7-fold higher than previously reported. Autotrophy was compared to ‘oxygen balanced’ mixotrophy where intracellular recirculation of O 2 and CO 2 take place. Aeration was not needed and 92% of the substrate carbon was converted into biomass. In mixotrophy biomass productivity was 1.8 times higher than autotrophic culture and linear growth was maintained at high biomass concentration (9.7 g x ·L −1 ). Light tolerance and high productivity in dense culture make our strain promising for mixotrophic outdoor cultivation. • Highest Galdieria sulphuraria autotrophic productivity (29 g·m −2 ·day −1 ) • Highest Galdieria sulphuraria mixotrophic productivity (51 g·m −2 ·day −1 ) • Oxygen balanced mixotrophy does not need aeration and 92% of substrate converted into biomass. • Galdieria sulphuraria can be cultivated at high light intensity. • In mixotrophy linear growth maintained at high biomass concentration (340 g x ·m −2 ).

Pilot scale production and evaluation of mechanical and thermal properties of P(3HB) from Bacillus megaterium cultivated on desugarized sugar beet molasses

AbstractProduction of poly[(R)‐3‐hydroxyalkanoate] (PHA) biopolyesters must become more cost efficient in order to be competitive with conventional plastics as well as other bioplastics. This study introduces a simple and reproducible large‐scale production process of poly(3‐hydroxybutyrate) P(3HB) with Bacillus megaterium based on desugarized sugar beet molasses and compares its material properties with commercially available types of PHA. A repeated batch cultivation system was evaluated in 1.2 L bioreactors and could be maintained for at least six cycles without a decrease in process stability, yielding a concentration of 20.4 g/L cell dry mass and 12 g/L P(3HB) per average process cycle. Feasibility of P(3HB) production was demonstrated at 500 L pilot scale to provide sufficient amounts of product for the determination of mechanical and thermal polymer properties. A volumetric productivity of 0.2 g L−1 h−1 was achieved. The resulting polymer exhibited a high tensile modulus of 3.73 GPa and tensile strength of 38.5 MPa. Despite a lower ductility and Vicat softening temperature, the properties of the recovered P(3HB) are comparable to commercial grade P(HB)HV polymers.

Fermentative Production of Fructo-Oligosaccharides Using Aureobasidium pullulans: Effect of Dissolved Oxygen Concentration and Fermentation Mode.

Fructo-oligosaccharides (FOS) are prebiotics with numerous health benefits. So far, the dissolved oxygen (DO) concentration control strategy for fermentative production of FOS is still unknown. In order to improve FOS production, the effects of DO concentration and fermentation mode on FOS using Aureobasidium pullulans were investigated in this study. The greatest FOS production (123.2 ± 6.2 g/L), with a yield of 61.6% ± 3.0% (g FOS/g sucrose), was obtained in batch culture under high DO concentration. Furthermore, repeated-batch culture revealed that enzyme production and FOS production were not closely associated with cell growth. By keeping the DO concentration above 5% in the repeated-batch culture, a maximum FOS concentration of 548.3 ± 37.4 g/L and yield of 68.6% ± 2.6% (g FOS/g sucrose) were obtained, which were 3.45% and 11.4% times higher than those obtained in the batch culture without DO control, respectively. Additionally, the ratios of 1-fructofuranosyl nystose (GF4) and 1,1,1,1-kestohexose (GF5) were 33.8% and 23.2%, respectively, in the product of repeated-batch culture, but these compounds were not detected in batch culture. Thus, it can be concluded that the DO concentration affects not only the yield of FOS but also the composition of FOS with different degrees of polymerization, which is the key factor in the fermentative production of FOS with a high polymerization degree.

Polymer fraction including exosomes derived from Chinese hamster ovary cells promoted their growth during serum-free repeated batch culture

While continuous (perfusion) culture of mammalian cells might reduce the reactor size owing to the high cell density, there is the problem of higher medium cost; however, this problem is expected to be solved by the reuse of growth-promoting components in the culture supernatant. The polymer fraction (PF, 10kDa-220nm) collected from the supernatant of serum-free repeated-batch culture of Chinese hamster ovary (CHO) cells in not only adhesion but also suspension promoted the cell growth in respective serum-free cultures. PF contained CD81-positive exosomes and proteins, both of which were necessary for its growth-promoting activity. Consequently, the medium cost for the continuous (perfusion) serum-free suspension culture of CHO cells may be decreased by the repeated collection and addition of PF that contains exosomes and growth factor proteins.

Alpha-Ketoglutaric Acid Production from a Mixture of Glycerol and Rapeseed Oil by Yarrowia lipolytica Using Different Substrate Feeding Strategies

The microbiological biosynthesis of α-ketoglutaric acid (KGA) has recently captured the attention of many scientists as an alternative to its common chemical synthesis. The present study aimed to evaluate the effect of the feeding strategy of substrates, i.e., glycerol (G = 20 g·dm−3) and rapeseed oil (O = 20 g·dm−3), on yeast growth and the parameters of KGA biosynthesis by a wild strain Yarrowia lipolytica A-8 in fed-batch and repeated-batch cultures. The effectiveness of KGA biosynthesis was demonstrated to depend on thiamine concentration and the substrate feeding method. In the fed-batch culture incubated with 3 µg·dm−3 of thiamine and a substrate feeding variant 2G(_OGO), KGA was produced in the amount of 62.1 g·dm−3 at the volumetric production rate of 0.37 g·dm−3·h−1. These values of KGA production parameters were higher than these obtained in the control culture (with rapeseed oil only). During 10 cycles of the 1788-h repeated-batch culture carried out acc. to the feeding strategy 2G(_OGO), in the last 5 cycles the yeast produced from 55.6 to 58.2 g·dm−3 of KGA and maximally 2.9 g·dm−3 of the pyruvic acid as a by-product.

Utilisation of tuna condensate waste from the canning industry as a novel substrate for polyhydroxyalkanoate production

Tuna condensate, an organic-rich by-product from the tuna canning industry, was assessed as a substrate for polyhydroxyalkanoate (PHA) production using Cupriavidus necator TISTR 1095. The effect of cultivation parameters on PHA accumulation was studied, including substrate concentration, carbon to nitrogen (C/N) ratio, initial pH-value control and fermentation strategies. For the bacterium, a biomass of 3.8 ± 0.1 g/L, PHA of 1.64 ± 0.1 g/L with PHA productivity of 0.027 g/L.h were obtained under batch cultivation using 100% tuna condensate with a C/N ratio of 88:1 and no control of pH. However, the PHA production was increased 1.3-fold when repeated-batch cultivation was applied. The highest biomass (7.5 ± 0.1 g/L) and PHA (3.8 ± 0.1 g/L) with 0.063 g/L.h of PHA productivity were achieved after the third cycle of repeated-batch cultivation. High chemical oxygen demand (COD) removal efficiency of 70% under the optimal condition was also demonstrated. The polymers generated by C. necator TISTR 1095 were characterised. The size of polymer granules was in the range of 0.7-0.8 μm. The polymer produced in the optimal medium under batch and repeated-batch cultivation was identified as poly(3-hydroxybutyrate-co-3-hydroxyvalerate) with a 20 mol% of 3-hydroxyvalerate. The molecular mass (Mn) and polydispersity of the polymer were 2 × 106 Da and 2.5, respectively. The results demonstrated that tuna condensate could be used as a cheap substrate for PHA production on an industrial scale.

Enhanced stability and nitrogen removal efficiency of Klebsiella sp. entrapped in chitosan beads applied in the domestic sewage system.

Although numerous denitrifying bacteria have been isolated and characterized, their capacity is seriously compromised by traditional inoculant addition and environmental stress in open bioreactors for wastewater treatment. In this study, a biocompatible material, chitosan, was used as a carrier to immobilize a simultaneously heterotrophic nitrifying–aerobic denitrifying bacterium Klebsiella sp., KSND, for continuous nitrogen removal from domestic wastewater in an open purification tank. The results showed that immobilization had no significant effect on cell viability and was beneficial for the reproduction and adhesion of cells. The entrapped KSND exhibited a slightly higher nitrogen removal efficiency of 90.09% than that of free KSND (87.69%). Subsequently, repeated batch cultivation experiments and analysis of the effects of organic contaminants and metal ions were performed using artificial wastewater and domestic wastewater. The findings revealed that the immobilized KSND beads presented desirable biophysical properties with good mechanical stability, cell viability, and enrichment, remarkable stability in organic contaminants and metal ions, and high efficiency nitrogen removal capacity. In conclusion, the developed immobilized denitrifying bacteria system has great potential for continuous wastewater treatment in open bioreactors.