Fermentation Technologies

Expression without boundaries: Cell-free protein synthesis in pharmaceutical research

by Marco G. Casteleijn

Co-authors: Arto Urtti, Sanjay Sarkhel. Published in "International Journal of Pharmaceutcs. 2012

Proteins are an increasingly important class of new drugs. Pharmaceutical proteins are usually expressed in cell based... more Proteins are an increasingly important class of new drugs. Pharmaceutical proteins are usually expressed in cell based systems in the development phase and in production, and although cell free methods have recently emerged they have not been used widely for therapeutic protein development or production. Cell free expression methodology is well suited for pharmaceutical protein expression and engineering and will probably become more commonly used in the future. Cell free expression allows protein engineering in high throughput format, flexible strategies for glycosylation and chemical conjugation, and allows easy use of unnatural amino acids as building blocks of proteins. Thus, cell free expression can be used to modify protein solubility, stability, and pharmacokinetics of therapeutic proteins. Likewise, it is potentially useful in protein development for biomaterial matrices, nanoparticles, and vaccines. This review illustrates the potential of cell free expression in pharmaceutical protein research and development while highlighting both advantages and limitations of the method.

Fermentation reactor coupled with capillary electrophoresis for on-line bioprocess monitoring

by Anne Menert

Tahkoniemi, H.; Helmja, K.; Menert, A.; Kaljurand, M.Journal of Pharmaceutical and Biomedical Analysis 41 (2006) 1585–1591

In this paper, a filter probe integrated into a computerized pneumatic sampler for capillary electroseparations was... more In this paper, a filter probe integrated into a computerized pneumatic sampler for capillary electroseparations was developed for an on-line monitoring of bioprocesses. The optimization of the performance of the coupled system was done by using a response surface modeling and three-level two-factor design. The resolution was found to be the most important parameter influencing the performance of an on-line analysis of the microbial metabolism. For the on-line analysis the migration time and detection limit were also found to be important parameters. Different parameters were combined by using an overall desirability function to find optimum conditions for all parameters. The equipment with an optimized separation protocol was used to monitor the bioaccumulation of Cu, Zn, Co and Cd (with detection limits 0.46, 0.37, 1.2, 0.84 mM correspondingly) by the Rhodococcus sp. bacteria isolated from the highly polluted technogenic soil of northeastern Estonia during a 2-week experiment.

Enzyme controlled glucose auto-delivery for high cell density cultivations in microplates and shake flasks

by Marco G. Casteleijn

High-cell density cultivation technique for microplates

by Marco G. Casteleijn

High-temperature cultivation and 5 [] mRNA optimization are key factors for the efficient overexpression of thermostable Deinococcus geothermalis purine nucleoside …

by Marco G. Casteleijn

Controlled high-cell density cultivation in shake flasks

by Marco G. Casteleijn

Utilization of palm kernel cake for production of β-glucosidase by Aspergillus niger FTCC 5003 in solid substrate fermentation using an aerated column bioreactor

by Peyman Abdeshahian

Abdeshahian, P., Samat, N & Wan Yusoff, W.M. 2010.Biotechnology. 9:17-24.

Xylanase production by Aspergillus niger FTCC 5003 using palm kernel cake in fermentative bioprocess

by Peyman Abdeshahian

Abdeshahian, P., Samat, N & Wan Yusoff, W.M. 2009.Pakistan Journal of Biological Sciences. 12: 1049-1055.

Production of β-xylosidase by Aspergillus niger FTCC 5003 using palm kernel cake in a packed-bed bioreactor

by Peyman Abdeshahian

Abdeshahian, P., Samat, N & Wan Yusoff, W.M. 2010.Journal of Applied
Sciences. 10: 419-424.

Solid substrate fermentation for cellulase production using palm kernel cake as a renewable lignocellulosic source in packed-bed bioreactor

by Peyman Abdeshahian

Abdeshahian, P., Samat, N., Abdul Hamid, A & Wan Yusoff, W.M.
2011.Biotechnology. Bioprocess. Engineering . 16: 238-
244.

Utilization of palm kernel cake for production of β-mannanase by Aspergillus niger FTCC 5003 in solid substrate fermentation using an aerated column bioreactor

by Peyman Abdeshahian

Abdeshahian, P., Samat, N., Abdul Hamid, A & Wan Yusoff, W.M.
2010.Journal of Industrial Microbiology &
Biotechnology 37: 103-109.

Nutrient Balance and Metabolic Analysis in a Kluyveromyces marxianus Fermentation with Lactose-Added Whey

by Javier Parrondo

Parrondo, J., García, L.A. and Díaz M. Nutrient Balance and Metabolic Analysis in a Kluyveromyces marxianus Fermentation with Lactose-Added Whey. Brazilian Journal of Chemical Engineering, 2009, 26(3) 445-456.

Addition of lactose on whey to produce an alcoholic product by fermentation is optimized in order
to maximise... more Addition of lactose on whey to produce an alcoholic product by fermentation is optimized in order
to maximise final ethanol concentrations and lactose consumption. The effect of the supplementation of the
broth with yeast extract, ammonium sulphate, oxygen, protein, peptides and the vitamins nicotinic acid,
biotin, pantothenic acid and inositol on aerobic cell growth was also studied. The Crabtree-negative yeast
Kluyveromyces marxianus is employed in this study, so oxygen should enhance cell growth and reduce
ethanol production. Addition of yeast extract, a source of vitamins, shifts metabolism towards fermentation.
The same effect is observed when nicotinic acid and biotin are added to the medium. Individual and mixed
effects of the four assayed vitamins are studied, showing that combinations of two or more vitamins
diminished cell growth and lactose consumption and increased ethanol production.

De la tecnología del medio ambiente de la destruccion de materia organica a la de su transformacion y aprovechamiento: suero lacteo (ii)

by Javier Parrondo

The road to sustainability implies obtaining the greatest benefits from the by-products generated during production... more The road to sustainability implies obtaining the greatest benefits from the by-products generated during production processes, in such a way that they are no longer considered waste as they have acquired added value. The specific characteristics of whey make it an excellent substrate for
biological transformation. A huge number of valuable products are obtained and the contaminant element of the by-product is
reduced . In this work, four possible transformations for whey by inoculation with specific microorganisms are analysed: production of alcoholic beverages/ ethanol, vinegar, enzymes and biomass.

Production of an alcoholic beverage by fermentation of whey permeate with kluyveromyces fragilis I: primary methabolism

by Javier Parrondo

J. Parrondo, L.A. Garcia and M. Diaz

Production of an alcoholic beverage by fermentation of whey permeate with kluyveromyces fragilis II: aroma composition

by Javier Parrondo

J. Parrondo, L. A. Garcia and M. Diaz

A Note – Production of Vinegar from Whey

by Javier Parrondo

Javier Parrondo, Mónica Herrero, Luis A. Garcia and Mario Díaz

Cheese whey supplemented with lactose was employed to produce vinegar. Whey was first transformed into an alcoholic... more Cheese whey supplemented with lactose was employed to produce vinegar. Whey was first transformed into an alcoholic beverage via fermentation with the yeast Kluyveromyces fragilis, and then the alcoholic product was employed as a substrate for an acetic acid fermentation. The bacteria used for the acetic acid production process were isolated from a starter culture provided by a vinegar factory. The bacteria employed were classified as Acetobacter pasteurianus. The vinegar obtained had a concentration of acetic acid between 5 and 6% (v/v). Ethyl acetate and fusel alcohols (isobutanol, 2-methyl-1-butanol and 3-methyl-1-butanol) were detected in the final product. The process and the substrates employed satisfied FAO requirements that the product was acceptable for human consumption. The efficiency of biotransformation
of ethanol into acetic acid was 84%.

A novel growth strategy for propagation and bacteriocin production of Lactobacilli

by Myrto-Panagiota Zacharof

Posted Presented in 8th World Congress of Chemical Engineering, Montreal Canada 2009

A novel growth strategy for propagation and bacteriocin production of Lactobacilli

M.P.Zacharof¹ and R.W.... more A novel growth strategy for propagation and bacteriocin production of Lactobacilli

M.P.Zacharof¹ and R.W. Lovitt²

Multidisciplinary Nanotechnology Center, Swansea University, Swansea, SA2 8PP, UK¹
School of Engineering, Multidisciplinary Nanotechnology Center, Swansea University, Swansea, SA2 8PP, UK²

Abstract
Lactobacilli belong to the group of lactic acid bacteria (LAB), widely used in the dairy industry nowadays. The bacteria are used as a natural acidifier for the inoculation of bulk quantities of milk and vegetables in order to produce a variety of fermented products. As such, large quantities of their biomass are necessary. Furthermore they produce natural antimicrobial substances during their growth, called bacteriocins. The possibility of producing these substances in mass quantities will be investigated through several techniques. The selected Lactobacilli, L.plantarum NCIMB 8014, L.casei NCIMB 11970 and L.lactis NCIMB 8586 were grown into simple batch cultures without pH control where their physicochemical needs were determined. Through the determination of the optimum nutritional conditions for the propagation of the Lactobacilli, an optimised medium for growth occurred.
The metabolism of the Lactobacilli was determined as homofermentative, mainly producing lactic acid. A simple liquid turbidometric method was developed to test the bacteriocin productivity of the selected bacteria. The efficiency and potency of the bacteriocin produced was tested against L.delbruckii subsp.lactis NCIMB 8117.
The optimised medium was further filtrated via membrane modules of 4 and 30 kDa and autoclaved. The medium’s efficiency in supporting the growth and the bacteriocin production of the bacilli was tested and comparative studies between filtrated and unfiltrated media were done.

Keywords: LAB, Bacteriocins, Target strain, Ultrafiltration, Nanofiltration, Growth rate, Doubling time

Development of an optimised growth strategy for intensive propagation, lactic acid and bacteriocin production of selected strains of Lactobacilli genus

by Myrto-Panagiota Zacharof

Published in International Journal of Chemical Engineering and Applications, Vol. 1, No. 1, June 2010 ISSN: 2010-0221

Abstract -
Lactobacilli belong to the group of lactic acid bacteria (LAB), widely used in the food industry... more Abstract -
Lactobacilli belong to the group of lactic acid bacteria (LAB), widely used in the food industry nowadays. These microorganisms have several distinguishing abilities such as the production of lactic acid, enzymes such as β-galactosidase and natural antimicrobial substances called bacteriocins. They are mainly used as a natural acidifier for the inoculation of bulk quantities of milk and vegetables in order to produce a variety of fermented products. As such, large quantities of their biomass and the end products of their metabolism are necessary. The possibility of producing these substances in mass quantities will be investigated through several techniques. The selected Lactobacilli, L.plantarum NCIMB 8014, L.casei NCIMB 11970, L.lactis NCIMB 8586were grown into simple batch cultures without pH control where their physicochemical needs were determined. Through the determination of the optimum nutritional conditions for the propagation of the Lactobacilli, an optimised medium for growth occurred. The optimum pH conditions for the growth of the bacilli were determined as well as parameters such as cellular yield coefficient, substrate and starter inoculums concentration and lactic acid rate and production. The metabolism of the Lactobacilli was determined as homofermentative, mainly producing lactic acid. The efficiency of the medium combining al the optimised parameters, enhancing the productivity biomass and therefore bacteriocins production from the strains, was tested on a 2L STR reactor operated batch wise with continuous pH control. A simple liquid turbidometric method was developed to test the bacteriocin productivity of the selected bacteria. The activity and potency of the bacteriocin produced was tested against L.delbruckii subsp.lactis NCIMB 8117.
Index Terms-Doubling time, Growth rate, Lactic acid, Nisin

Optimization of growth conditions for intensive propagation, growth development and lactic acid production of selected strains of Lactobacilli

by Myrto-Panagiota Zacharof

Paper Presented in Chemeca 2009 27–30 September 2009 Burswood Entertainment Complex, Perth, Australia

Optimization of growth conditions for intensive propagation, growth development and lactic acid production of selected... more Optimization of growth conditions for intensive propagation, growth development and lactic acid production of selected strains of Lactobacilli

M.P.Zacharof¹, R.W. Lovitt² and K. Ratanapongleka ³

Multidisciplinary Nanotechnology Center, Swansea University, Swansea, SA2 8PP, UK¹
School of Engineering, Multidisciplinary Nanotechnology Center, Swansea University, Swansea, SA2 8PP, UK²
Department of Chemical Engineering, Faculty of Engineering, Ubon Rajathanee University, Ubon Rajathanee 34190, Thailand³


Abstract
Lactobacilli belong to the group of lactic acid bacteria (LAB), widely used in the food industry These microorganisms have several distinguishing abilities such as the production of lactic acid, enzymes such as β-galactosidase and natural antimicrobial substances called bacteriocins. They are mainly used as a natural acidifier for the inoculation of bulk quantities of milk and vegetables in order to produce a variety of fermented products. As such, large quantities of their biomass and the end products of their metabolism are necessary. The possibility of producing these substances in mass quantities was investigated through several techniques. Selected Lactobacilli, L.plantarum NCIMB 8014, L.casei NCIMB 11970, L.lactis NCIMB 8586 and L.delbruckii NCIMB 11778 were grown into batch cultures without pH control where their physicochemical needs were determined. Through the determination of the optimum nutritional conditions for the propagation of the Lactobacilli, an optimised medium for growth was developed. The efficiency of the medium was tested in a 2L STR reactor operated batch wise with continuous pH control. The optimum pH conditions for the growth of the bacilli were determined as well as parameters such as cellular yield coefficient, substrate and starter inoculum concentration and lactic acid rate and production. The metabolism of the Lactobacilli was determined as homofermentative, mainly producing lactate. Efficiency of the optimized medium was evaluated in terms of growth rate and doubling time through the spectrophotometric measurement of cellular biomass. The medium strongly supports the growth of the Lactobacilli giving a growth rate between 0.22 to 0.32 h-1

Keywords: LAB, STR, Growth rate, Doubling time, lactic acid, nutrient medium

The importance of Lactobacilli in contemporary food and pharmaceutical industry A review article

by Myrto-Panagiota Zacharof

Proceedings of 2010 International Conference on Chemical Engineering and Applications (CCEA 2010) Singapore, 26-28 February, 2010 ISBN13: 978-1-84626-023-0

Abstract
Fermentation technology has been a widely researched and exploited field of the science of... more Abstract
Fermentation technology has been a widely researched and exploited field of the science of biotechnology. Through out the recent years the vast majority of microbial groups have been tested for the production of beneficial compounds especially for the replacement of products produced by petrol such as lactic acid. A bacterial group that heavily attracts attention due to its products are Lactic Acid Bacteria (LAB) and especially Lactobacilli. Lactobacilli are widely used in the food and pharmaceutical industry nowadays. These microorganisms have several distinguishing features based on their main ability to ferment carbohydrates such as the production of acids, enzymes and natural antimicrobial substances called bacteriocins. They are mainly used as natural acidifiers for the inoculation of bulk quantities of milk and vegetables in order to produce a variety of fermented products. As such, large quantities of their biomass and the end products of their metabolism are necessary. In this article some of the most important uses of Lactobacilli in the industry will be reviewed. Emphasis will be given in the production of lactic acid, β-galactosidase and lantibiotics through the usage of modern fermentation technology.
Keywords: LAB, fermentation technology, food industry, β-galactosidase, lactic acid,