Saúl Alonso

Efficient lactobionic acid production from whey by Pseudomonas taetrolens under pH-shift conditions.

Lactobionic acid finds applications in the fields of pharmaceuticals, cosmetics and medicine. The production of lactobionic acid from whey by Pseudomonas taetrolens was studied in shake-flasks and in a bioreactor. Shake-flask experiments showed that lactobionic acid was a non-growth associated product. A two-stage pH-shift bioconversion strategy with a pH-uncontrolled above 6.5 during the growth phase and maintained at 6.5 during cumulative production was adopted in bioreactor batch cultures. An inoculation level of 30% promoted high cell culture densities that triggered lactobionic acid production at a rate of 1.12 g/Lh. This methodology displayed efficient bioconversion with cheese whey as an inexpensive substrate for lactobionic acid production.

Bio-production of lactobionic acid: Current status, applications and future prospects

Lactobionic acid has appeared on the commercial scene as a versatile polyhydroxy acid with numerous promising applications in the food, medicine, pharmaceutical, cosmetics and chemical industries. This high value-added bio-product has recently received growing attention as a bioactive compound, providing an excellent chemical platform for the synthesis of novel potentially biocompatible and biodegradable drug delivery vehicles. Recent advances in tissue engineering and nanomedicine have also underlined the increased importance of this organic acid as a key biofunctionalization agent. The growing commercial relevance of lactobionic acid has therefore prompted the development of novel systems for its biotechnological production that are both sustainable and efficient. The present review explores recent advances and studies related to lactobionic acid bio-production, whether through microbial or enzymatic approaches, highlighting the key bioprocessing conditions for enhanced bio-production. Detailed overviews of the current microbial cell factories as well as downstream processing methodologies for lactobionic acid production are also presented. Furthermore, the potential prospects and current applications of this polyhydroxy acid are also discussed, with an emphasis on the role of lactobionic acid as a key platform in the development of novel drugs, biomaterials, nanoparticles and biopolymer systems.

Microbial production of specialty organic acids from renewable and waste materials

Abstract Microbial production of organic acids has become a fast-moving field due to the increasing role of these compounds as platform chemicals. In recent years, the portfolio of specialty fermentation-derived carboxylic acids has increased considerably, including the production of glyceric, glucaric, succinic, butyric, xylonic, fumaric, malic, itaconic, lactobionic, propionic and adipic acid through innovative fermentation strategies. This review summarizes recent trends in the use of novel microbial platforms as well as renewable and waste materials for efficient and cost-effective bio-based production of emerging high-value organic acids. Advances in the development of robust and efficient microbial bioprocesses for producing carboxylic acids from low-cost feedstocks are also discussed. The industrial market scenario is also reviewed, including the latest information on the stage of development for producing these emerging bio-products via large-scale fermentation.

Role of dissolved oxygen availability on lactobionic acid production from whey by Pseudomonas taetrolens.

The influence of dissolved oxygen availability on cell growth and lactobionic acid production from whey by Pseudomonas taetrolens has been investigated for the first time. Results from pH-shift bioreactor cultivations have shown that high agitation rate schemes stimulated cell growth, increased pH-shift values and the oxygen uptake rate by cells, whereas lactobionic acid production was negatively affected. Conversely, higher aeration rates than 1.5 Lpm neither stimulated cell growth nor lactobionic acid production (22% lower for an aeration rate of 2 Lpm). Overall insights into bioprocess performance enabled the implementation of 350 rpm as the optimal agitation strategy during cultivation, which increased lactobionic productivity 1.2-fold (0.58-0.7 g/Lh) compared to that achieved at 1000 rpm. Oxygen supply has been shown to be a key bioprocess parameter for enhanced overall efficiency of the system, representing essential information for the implementation of lactobionic acid production at a large scale.

Feeding strategies for enhanced lactobionic acid production from whey by Pseudomonas taetrolens

High-level production of lactobionic acid from whey by Pseudomonas taetrolens under fed-batch fermentation was achieved in this study. Different feeding strategies were evaluated according to the physiological status and fermentation performance of P. taetrolens. A lactobionic acid titer of 164 g/L was obtained under co-feeding conditions affording specific and volumetric productivities of 1.4 g/g h and 2.05 g/L h, respectively. Flow cytometry assessment revealed that P. taetrolens cells exhibited a robust physiological status, which makes them particularly well-suited for employing concentrated nutrient solutions to further prolong the growth and production phases. Such detailed knowledge of the physiological status has been revealed to be a key issue to further support the development of high-yield lactobionic acid production processes under feeding strategies. The present study has demonstrated the feasibility of P. taetrolens to achieve high-level bio-production of lactobionic acid from whey through fed-batch cultivation, suggesting its major potential for industrial-scale implementation.

Simultaneous production of lactobionic and gluconic acid in cheese whey/glucose co-fermentation by Pseudomonas taetrolens

Substrate versatility of Pseudomonas taetrolens was evaluated for the first time in a co-fermentation system combining cheese whey and glucose, glycerol or lactose as co-substrates. Results showed that P. taetrolens displayed different production patterns depending on the co-substrate supplied. Whereas the presence of glucose led to a simultaneous co-production of lactobionic (78g/L) and gluconic acid (8.8g/L), lactose feeding stimulated the overproduction of lactobionic acid from whey with a high specific productivity (1.4g/gh) and yield (100%). Co-substrate supply of glycerol conversely led to reduced lactobionic acid yield (82%) but higher cell densities (1.8g/L), channelling the carbon source towards cell growth and maintenance. Higher carbon availability impaired the metabolic activity as well as membrane integrity, whereas lactose feeding improved the cellular functionality of P. taetrolens. Insights into these mixed carbon source strategies open up the possibility of co-producing lactobionic and gluconic acid into an integrated single-cell biorefinery.

Selection method of pH conditions to establish Pseudomonas taetrolens physiological states and lactobionic acid production.

Microbial physiological responses resulting from inappropriate bioprocessing conditions may have a marked impact on process performance within any fermentation system. The influence of different pH-control strategies on physiological status, microbial growth and lactobionic acid production from whey by Pseudomonas taetrolens during bioreactor cultivations has been investigated for the first time in this work. Both cellular behaviour and bioconversion efficiency from P. taetrolens were found to be negatively influenced by pH-control modes carried out at values lower than 6.0 and higher than 7.0. Production schemes were also influenced by the operational pH employed, with asynchronous production from damaged and metabolically active subpopulations at pH values lower than 6.0. Moreover, P. taetrolens showed reduced cellular proliferation and a subsequent delay in the onset of the production phase under acidic conditions (pH < 6.0). Unlike cultivations performed at 6.5, both pH-shift and pH-stat cultivation strategies performed at pH values lower than 6.0 resulted in decreased lactobionic acid production. Whereas the cellular response showed a stress-induced physiological response under acidic conditions, healthy functional cells were predominant at medium operational pH values (6.5–7.0). P. taetrolens thus displayed a robust physiological status at initial pH value of 6.5, resulting in an enhanced bioconversion yield and lactobionic acid productivity (7- and 4-fold higher compared to those attained at initial pH values of 4.5 and 5.0, respectively). These results have shown that pH-control modes strongly affected both the physiological response of cells and the biological performance of P. taetrolens, providing key information for bio-production of lactobionic acid on an industrial scale.

Flow cytometric characterization of bacterial abundance and physiological status in a nitrifying-denitrifying activated sludge system treating landfill leachate

Flow cytometry has recently been presented as a research tool in the assessment of the viability/activity of activated sludge from municipal wastewater treatment plants, but it has not put in practice for industrial biotreatments yet. In this study, for the first time ever, the reliability and significance of the multiparameter flow cytometry applied to the biological nitrification-denitrification treatment of leachate have been evaluated. Using a double staining procedure (cFDA/PI), the viable, damaged, and dead subpopulations were determined, and the results were compared to those obtained with conventional methods, such as nitrogen and oxygen uptake rates or plate counting. Flow cytometry showed that viable cells represented approximately 47% of the total population, whereas active cells accounted for 90%. For both sludge from nitrification and denitrification processes, with less than 1% of them being also culturable in plate. Either flow cytometry or uptake rates revealed that health status of sludge remained constant throughout the biotreatment, which is consistent with the high recirculation rates. Under anaerobic starvation conditions, physiological status of sludge remained constant as well as specific oxygen and denitrification rates. Nevertheless, both the culturability in plate and the nitrification rate significantly decreased. These findings proved that multiparameter flow cytometry is a useful tool for the assessment of the viability and activity of sludge from a nitrification-denitrification biotreatment process. These results gathered all the bacterial communities in the sludge, so the decay in minority populations, such as nitrifying bacteria, requires the use of a complementary technique to evaluate specific activities.