Dihydroxyacetone production via heterogeneous biotransformations of crude glycerol.

In this work, several immobilization strategies for Gluconobacter oxydans NBRC 14819 (Gox) were tested in the bioconversion of crude glycerol to dihydroxyacetone (DHA). Agar, agarose and polyacrylamide were evaluated as immobilization matrixes. Glutaraldehyde crosslinked versions of the agar and agarose preparations were also tested.

Green Production of Cladribine by Using Immobilized -Deoxyribosyltransferase from Stabilized through a Double Covalent/Entrapment Technology.

Nowadays, enzyme-mediated processes offer an eco-friendly and efficient alternative to the traditional multistep and environmentally harmful chemical processes. Herein we report the enzymatic synthesis of cladribine by a novel 2'-deoxyribosyltransferase (NDT)-based combined biocatalyst. To this end, NDT (NDT) was successfully immobilized through a two-step immobilization methodology, including a covalent immobilization onto glutaraldehyde-activated biomimetic silica nanoparticles followed by biocatalyst entrapment in calcium alginate.

Decitabine bioproduction using a biocatalyst with improved stability by adding nanocomposites.

A novel IDA-LaNDT derivative was able to reach the highest productivity in the biosynthesis of a well-known antitumoral agent called decitabine. However, the combination of two simple and inexpensive techniques such as ionic absorption and gel entrapment with the incorporation of a bionanocomposite such as bentonite significantly improved the stability of this biocatalyst. These modifications allowed the enhancement of storage stability (for at least 18 months), reusability (400 h of successive batches without significant loss of its initial activity), and thermal and solvent stability with respect to the non-entrapped derivative.

Biotransformation of cladribine by a nanostabilized extremophilic biocatalyst.

Cladribine (2-chloro-2'-deoxy-β-d-adenosine) is a 2'-deoxyadenosine analogue, approved by the FDA for the treatment of hairy cell leukemia and more recently has been proved for therapeutic against many autoimmune diseases as multiple sclerosis. The biosynthesis of this compound using Thermomonospora alba CECT 3324 as biocatalyst is herein reported. This thermophilic microorganism was successfully entrapped in polyacrylamide gel supplemented with nanoclays such as bentonite.

Hyperstabilization of a thermophile bacterial laccase and its application for industrial dyes degradation.

In the present study, a novel extracellular laccase isolated from ATCC 10149 was entrapped in a bionanocomposite matrix consisting of copper alginate (Cu-alginate) supplemented with the nanoclay bentonite. After optimization, this nanobiocatalyst was able to degrade up to 90% of Remazol Brilliant Blue R (RBBR) without the addition of redox mediators and retained 70% of its initial activity for at least 1440 h, equivalent to more than 288 uses. The incorporation of nanoclay allowed alginate beads to be used in alkaline pH and strengthened its mechanical properties.

Biotransformation of cladribine by a magnetic immobilizated biocatalyst of Lactobacillus animalis.

A stable biocatalyst with magnetic properties based on immobilized Lactobacillus animalis ATCC 35,046 to obtain 2-chloroadenine-2'-deoxyriboside, known as cladribine, is reported for the first time. This nucleoside analogue is an antitumor agent used in the treatment of a wide variety of types of leukemia. In this study, an eco-compatible and alternative bioprocess to obtain cladribine was developed.

Whole Cell Entrapment Techniques.

Microbial whole cells are efficient, ecological, and low-cost catalysts that have been successfully applied in the pharmaceutical, environmental, and alimentary industries, among others.Microorganism immobilization is a good way to carry out the bioprocess under preparative conditions. The main advantages of this methodology lie in their high operational stability, easy upstream separation, and bioprocess scale-up feasibility.

Biotransformation of cladribine using a stabilized biocatalyst in calcium alginate beads.

Cladribine is a nucleoside analogue widely used in the pharmaceutical industry for the treatment of several neoplasms, including hairy-cell leukemia among others. This compound has also shown efficacy in the treatment of autoimmune diseases such as rheumatoid arthritis and multiple sclerosis. In this work, a green bioprocess for cladribine biosynthesis using immobilized Arthrobacter oxydans was developed.

Development of a high efficient biocatalyst by oriented covalent immobilization of a novel recombinant 2'-N-deoxyribosyltransferase from Lactobacillus animalis.

The 2'-N-deoxyribosyltransferases [NDT; EC 2.4.2.

Immobilization techniques applied to the development of biocatalysts for the synthesis of nucleoside analogue derivatives.

Background: Nucleoside analogue (NAs) derivatives comprise a large family of pharmaceuticals clinically used as antitumoral and antiviral compounds. Originally, the production of NAs involved chemical synthesis, but a greener bioproduction alternative exists and involves the use of enzymes that catalyze transglycosylation reactions between modified purinic or pyrimidinic bases and sugars. To be considered as an option for industrial application, it is vital to immobilize these biocatalysts.

Saccharification of citrus wastes by immobilized polygalacturonase in an improved alginate matrix.

Enzyme immobilization using hydrogels is a low-cost and effective system for the degradation of bulk pectin derived from orange industry residues. Polygalacturonases obtained from four different bacterial strains of genus were immobilized in alginate gel and assayed for pectin hydrolysis. The enzyme from ATCC 10897 proved to be superior and more stable within the alginate matrix.

Microbiological and serological control of Escherichia coli O157: H7 in kindergarten staff in Buenos Aires city and suburban areas.

Shiga toxin (Stx)-producing Escherichia coli (STEC) infections are implicated in the development of the life-threatening hemolytic-uremic syndrome (HUS). Despite the magnitude of the social and economic problems caused by HUS, no licensed vaccine or effective therapy is currently available for human use. Prevention of STEC infections continues being the most important measure to reduce HUS incidence.

Biosynthesis of an antiviral compound using a stabilized phosphopentomutase by multipoint covalent immobilization.

Ribavirin is a synthetic guanosine analogue with a broad-spectrum of antiviral activity. It is clinically effective against several viruses, such as respiratory syncytial virus, several hemorrhagic fever viruses and HCV when combined with pegylated interferon-α. Phosphopentomutase (PPM) catalyzes the transfer of intramolecular phosphate (from C1 to C5) on ribose, and is involved in pentose phosphate pathway and in purine metabolism.

Stabilization by multipoint covalent attachment of a biocatalyst with polygalacturonase activity used for juice clarification.

Derivatized-agarose supports are suitable for enzyme immobilization by different methods, taking advantage of different physical, chemical and biological conditions of the protein and the support. In this study, agarose particles were modified with MANAE, PEI and glyoxyl groups and evaluated to stabilize polygalacturonase from Streptomyces halstedii ATCC 10897. A new immobilized biocatalyst was developed using glyoxyl-agarose as support; it exhibited high performance in degrading polygalacturonic acid and releasing oligogalacturonides.

Bioproduction of ribavirin by green microbial biotransformation.

Ribavirin is an antiviral compound widely used in Hepatitis C Virus therapy. Biotransformation of this nucleoside analogue using ATCC 12407 as biocatalyst is herein reported. Reaction parameters such as microorganism amounts, substrate ratio and temperature were optimized reaching conversion yields of 86%.

Whole cell entrapment techniques.

Microbial whole cells are efficient, ecological, and low-cost catalysts that have been successfully applied in the pharmaceutical, environmental, and alimentary industries, among others. Microorganism immobilization is a good way to carry out the bioprocess under preparative conditions. The main advantages of this methodology lie in their high operational stability, easy upstream separation and bioprocess scale-up feasibility.

Biosynthesis of anti-HCV compounds using thermophilic microorganisms.

This work describes the application of thermophilic microorganisms for obtaining 6-halogenated purine nucleosides. Biosynthesis of 6-chloropurine-2'-deoxyriboside and 6-chloropurine riboside was achieved by Geobacillus stearothermophilus CECT 43 with a conversion of 90% and 68%, respectively. Furthermore, the selected microorganism was satisfactorily stabilized by immobilization in an agarose matrix.

Biotransformation of 2,6-diaminopurine nucleosides by immobilized Geobacillus stearothermophilus.

An efficient and green bioprocess to obtain 2,6-diaminopurine nucleosides using thermophilic bacteria is herein reported. Geobacillus stearothermophilus CECT 43 showed a conversion rate of 90 and 83% at 2 h to obtain 2,6-diaminopurine-2'-deoxyriboside and 2,6-diaminopurine riboside, respectively. The selected biocatalyst was successfully stabilized in an agarose matrix and used to produce up to 23.

Green biosynthesis of floxuridine by immobilized microorganisms.

This work describes an efficient, simple, and green bioprocess for obtaining 5-halogenated pyrimidine nucleosides from thymidine by transglycosylation using whole cells. Biosynthesis of 5-fluoro-2'-deoxyuridine (floxuridine) was achieved by free and immobilized Aeromonas salmonicida ATCC 27013 with an 80% and 65% conversion occurring in 1 h, respectively. The immobilized biocatalyst was stable for more than 4 months in storage conditions (4 °C) and could be reused at least 30 times without loss of its activity.

Immobilized Escherichia coli BL21 as a catalyst for the synthesis of adenine and hypoxanthine nucleosides.

Different supports, such as alginate, agar, agarose, and polyacrylamide, were used to immobilize Escherichia coli BL 21 by entrapment techniques. The transglycosylation reaction involved in the synthesis of adenosine from uridine and adenine was chosen as a model system to study the characteristics of these biocatalysts. Whole cells immobilized on agarose proved to be optimal and could be used up to 30 times without significant loss of activity.