Alcohol dehydrogenase stabilization by additives under industrially relevant reaction conditions

Spickermann, Dominik; Kara, Selin; Barackov, Ivana; Hollmann, Frank; Schwaneberg, Ulrich; Duenkelmann, Pascal; Leggewie, Christian

doi:10.1016/j.molcatb.2013.11.015

Cited by 20 publications

(8 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As such, it represents a proof of concept with absolutely considerable potential for further improvements. Future process development studies for enhancing productivity14 will focus on 1) optimisation of biocatalyst concentrations, ratios, etc., 2) the use of high substrate concentrations for achieving enzyme‐to‐substrate ratios >50 (g g −1 )15 by biphasic and water‐free systems, 3) stabilisation of enzymes by means of immobilisation16 or using additives 17. Special attention will be devoted to in situ removal of the lactone product and to develop an integrated ECL polymerisation under controlled conditions.…”

Section: Methodsmentioning

confidence: 99%

A Bi‐enzymatic Convergent Cascade for ε‐Caprolactone Synthesis Employing 1,6‐Hexanediol as a ‘Double‐Smart Cosubstrate’

et al. 2015

Self Cite

View full text Add to dashboard Cite

A bi‐enzymatic cascade consisting of a Baeyer–Villiger monooxygenase and an alcohol dehydrogenase (ADH) was designed in a convergent fashion to utilise two molar equivalents of cyclohexanone (CHO) and one equivalent of 1,6‐hexanediol as a ‘double‐smart cosubstrate’ to produce ε‐caprolactone (ECL) with water as sole by‐product. The convergent enzymatic cascade reaction reported herein, is performed at ambient conditions in water, is self‐sufficient with respect to cofactor, and incorporates all starting materials into the desired product, ECL. Among different enzymes explored, the reaction catalysed by cyclohexanone monooxygenase from Acinetobacter sp. NCIMB 9871 coupled with ADH from Thermoanaerobacter ethanolicus showed the best results, reaching 91 % conversion of CHO after 24 h with a product titre of 2 g L−1. Scale‐up of the coupled system (50 mL) performed better than the small‐scale reactions and >99 % conversion of CHO and ECL concentration of 20 mM were achieved within 18 h.

show abstract

Section: Methodsmentioning

confidence: 99%

A Bi‐enzymatic Convergent Cascade for ε‐Caprolactone Synthesis Employing 1,6‐Hexanediol as a ‘Double‐Smart Cosubstrate’

et al. 2015

Self Cite

View full text Add to dashboard Cite

show abstract

“…As example, enzymes applied in feed must be thermostable (due to the pelleting process) and must be stable or active at low pH (stomach of animals) (Viader‐Salvadó et al ., ). Also, additives such as salts to a high concentration can be used as additives for enzyme stabilization under industrially relevant conditions, and therefore the halophilic enzymes, such as alcohol dehydrogenases, may be desired for certain applications (Spickermann et al ., ). Clearly, novel backbones from metagenomes might meet these requirements.…”

Section: Final Considerations: Backbones Of Interest For Finding Markmentioning

confidence: 97%

“…They included the following: (i) harsh and broad reaction conditions such as a high substrate load (necessary to reduce the costs to be competitive), broad range of temperatures (at least should be stable at room temperature for a period of time as also storage might be an additional issue – think in detergent enzymes applied in warmer countries), broad range of pHs, water‐deficient reaction conditions, very high solvent concentrations (which for example might be necessary for subsequent downstream processing) and process stability (e.g. active for 12–24 h) (Spickermann et al ., ; Zuhse et al ., ); and (ii) the high stereoselectivity and high turnover rates (Singh, ). As example, enzymes applied in feed must be thermostable (due to the pelleting process) and must be stable or active at low pH (stomach of animals) (Viader‐Salvadó et al ., ).…”

Section: Final Considerations: Backbones Of Interest For Finding Markmentioning

confidence: 98%

Estimating the success of enzyme bioprospecting through metagenomics: current status and future trends

Ferrer

Martínez-Martínez

Bargiela

et al. 2015

Microbial Biotechnology

190

149

View full text Add to dashboard Cite

SummaryRecent reports have suggested that the establishment of industrially relevant enzyme collections from environmental genomes has become a routine procedure. Across the studies assessed, a mean number of approximately 44 active clones were obtained in an average size of approximately 53 000 clones tested using naïve screening protocols. This number could be significantly increased in shorter times when novel metagenome enzyme sequences obtained by direct sequencing are selected and subjected to high‐throughput expression for subsequent production and characterization. The pre‐screening of clone libraries by naïve screens followed by the pyrosequencing of the inserts allowed for a 106‐fold increase in the success rate of identifying genes encoding enzymes of interest. However, a much longer time, usually on the order of years, is needed from the time of enzyme identification to the establishment of an industrial process. If the hit frequency for the identification of enzymes performing at high turnover rates under real application conditions could be increased while still covering a high natural diversity, the very expensive and time‐consuming enzyme optimization phase would likely be significantly shortened. At this point, it is important to review the current knowledge about the success of fine‐tuned naïve‐ and sequence‐based screening protocols for enzyme selection and to describe the environments worldwide that have already been subjected to enzyme screen programmes through metagenomic tools. Here, we provide such estimations and suggest the current challenges and future actions needed before environmental enzymes can be successfully introduced into the market.

show abstract

“…These include: i) the ability to maintain activity and stability under harsh reaction conditions and a broad range of temperatures (enzymes should at least be stable at room temperatures for a significant period of time -storage might be an additional issue, which can refer to detergent enzymes applied in warmer countries); and ii) to be active and stable under a broad range of water-deficient reaction conditions and under process conditions (e.g., active for 12-24 h) (Viader-Galván et al 2010;Spickermann et al 2014;Zuhse et al 2015). Accordingly, the optimal temperature for activity and stability in, and resistance against, media containing organic solvents, are important parameters to examine.…”

Section: Hydrolytic Activities Of Est-lip Under Different Conditionsmentioning

confidence: 99%

Biodiversity for biocatalysis: A review of the α/β-hydrolase fold superfamily of esterases-lipases discovered in metagenomes

Ferrer

Bargiela

Martínez-Martínez

et al. 2015

Biocatalysis and Biotransformation

View full text Add to dashboard Cite

Natural biodiversity undoubtedly inspires biocatalysis research and innovation.Biotransformations of interest also inspire the search for appropriate biocatalysts in nature.Indeed, natural genetic resources have been found to support the hydrolysis and synthesis of not only common but also unusual synthetic scaffolds. The emerging tool of metagenomics has the advantage of allowing straightforward identification of activity directly applicable as biocatalysis. However, new enzymes must not only have outstanding properties in terms of performance but also other properties superior to those of well-established commercial preparations in order to successfully replace the latter. Esterases (EST) and lipases (LIP) from the α/ -hydrolase fold superfamily are among the enzymes primarily used in biocatalysis. Accordingly, they have been extensively examined with metagenomics. Here we provided an updated (October 2015) overview of sequence and functional datasets of 288 EST-LIP enzymes with validated functions that have been isolated in metagenomes and (mostly partially) characterized. Through sequence, biochemical and reactivity analyses we attempted to understand the phenomenon of variability and versatility within this group of enzymes and to implement this knowledge to identify sequences encoding EST-LIP which may be useful for biocatalysis. We found that the diversity of described EST-LIP polypeptides was not dominated by a particular type of protein or highly similar clusters of proteins but rather by diverse nonredundant sequences. Purified EST-LIP exhibited a wide temperature activity range of 10-85°C, although a preferred bias for a mesophilic temperature range (35-40ºC) was observed. At least 60% of the total characterized metagenomics-derived EST-LIP showed outstanding properties in terms of stability (solvent tolerance) and reactivity (selectivity and substrate profile), which are the features of interest in biocatalysis. We hope that, in the future, the search for and utilization of sequences similar to those already encoded and characterized EST-LIP enzymes 3 from metagenomes may be of interest for promoting unresolved biotransformations in the chemical industry. Some examples are discussed in this review.4

show abstract

Alcohol dehydrogenase stabilization by additives under industrially relevant reaction conditions

Cited by 20 publications

References 23 publications

A Bi‐enzymatic Convergent Cascade for ε‐Caprolactone Synthesis Employing 1,6‐Hexanediol as a ‘Double‐Smart Cosubstrate’

A Bi‐enzymatic Convergent Cascade for ε‐Caprolactone Synthesis Employing 1,6‐Hexanediol as a ‘Double‐Smart Cosubstrate’

Estimating the success of enzyme bioprospecting through metagenomics: current status and future trends

Biodiversity for biocatalysis: A review of the α/β-hydrolase fold superfamily of esterases-lipases discovered in metagenomes

Contact Info

Product

Resources

About