Improvement of thermostable aldehyde dehydrogenase by directed evolution for application in Synthetic Cascade Biomanufacturing

Steffler, Fabian; Guterl, Jan‐Karl; Sieber, Volker

doi:10.1016/j.enzmictec.2013.07.002

Cited by 33 publications

(25 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…“Heat” purification was also utilized during efforts to engineer a thermotolerant aldehyde dehydrogenase for glucose‐to‐ethanol CFME [49]. Here, reaction velocity and NAD + affinity were improved via protein refolding [84] and evolution by random mutagenesis [85]. Shorter pathways have also leveraged “heat” purification in conjunction with ammonium sulfate precipitation, a technique where the salt concentration is increased until proteins begin sequentially precipitating [86].…”

Section: Challenges and Opportunities In Cfmementioning

confidence: 99%

Cell‐free metabolic engineering: Biomanufacturing beyond the cell

Dudley

Karim

Jewett

2014

Biotechnology Journal

285

258

View full text Add to dashboard Cite

Industrial biotechnology and microbial metabolic engineering are poised to help meet the growing demand for sustainable, low-cost commodity chemicals and natural products, yet the fraction of biochemicals amenable to commercial production remains limited. Common problems afflicting the current state-of-the-art include low volumetric productivities, build-up of toxic intermediates or products, and byproduct losses via competing pathways. To overcome these limitations, cell-free metabolic engineering (CFME) is expanding the scope of the traditional bioengineering model by using in vitro ensembles of catalytic proteins prepared from purified enzymes or crude lysates of cells for the production of target products. In recent years, the unprecedented level of control and freedom of design, relative to in vivo systems, has inspired the development of engineering foundations for cell-free systems. These efforts have led to activation of long enzymatic pathways (>8 enzymes), near theoretical conversion yields, productivities greater than 100 mg L−1 hr−1, reaction scales of >100L, and new directions in protein purification, spatial organization and enzyme stability. In the coming years, CFME will offer exciting opportunities to (i) debug and optimize biosynthetic pathways, (ii) carry out design-build-test iterations without re-engineering organisms, and (iii) perform molecular transformations when bioconversion yields, productivities, or cellular toxicity limit commercial feasibility.

show abstract

Section: Challenges and Opportunities In Cfmementioning

confidence: 99%

Cell‐free metabolic engineering: Biomanufacturing beyond the cell

Dudley

Karim

Jewett

2014

Biotechnology Journal

285

258

View full text Add to dashboard Cite

show abstract

“…The performance of a module depends greatly on the efficiency of enzymes; therefore, increasing the stability, solubility, and catalytic efficiency of enzymes by engineering will markedly improve module performance. In fact, thermostable enzymes have been intensively used in modules because of their high operational stability [13] , [59] , [60] . In addition, the activity of several enzymes has been altered using enzyme engineering, allowing for the design of artificial pathways [11] , [28] , [61] .…”

Section: Discussionmentioning

confidence: 99%

Modules for in vitro metabolic engineering: Pathway assembly for bio-based production of value-added chemicals

Taniguchi

Okano

Honda

2017

Synthetic and Systems Biotechnology

View full text Add to dashboard Cite

Bio-based chemical production has drawn attention regarding the realization of a sustainable society. In vitro metabolic engineering is one of the methods used for the bio-based production of value-added chemicals. This method involves the reconstitution of natural or artificial metabolic pathways by assembling purified/semi-purified enzymes in vitro. Enzymes from distinct sources can be combined to construct desired reaction cascades with fewer biological constraints in one vessel, enabling easier pathway design with high modularity. Multiple modules have been designed, built, tested, and improved by different groups for different purpose. In this review, we focus on these in vitro metabolic engineering modules, especially focusing on the carbon metabolism, and present an overview of input modules, output modules, and other modules related to cofactor management.

show abstract

“…An efficient system needs to be developed whereby the pH of SPJ is raised without adding large amounts of base and where the nutrients from SPJ are preserved for subsequent biotechnological processes. enzymes from all domains of life, and individually engineered to meet the requirements of the cascade, can be combined to form an optimized pathway (Heinzelman et al, 2009;Lutz, 2010;Steffler et al, 2013). For the development of a cell-free system, that selectively deacidifies SPJ by reducing the amount of organic acids while producing a value-added compound, lactic acid is a suitable target substrate.…”

Section: Introductionmentioning

confidence: 99%

Deacidification of grass silage press juice by continuous production of acetoin from its lactate via an immobilized enzymatic reaction cascade

Hohagen

Schwarz

Schenk

et al. 2017

Bioresource Technology

Self Cite

View full text Add to dashboard Cite

An immobilized enzymatic reaction cascade was designed and optimized for the deacidification of grass silage press juice (SPJ), thus facilitating the production of bio-based chemicals. The cascade involves a three-step process using four enzymes immobilized in a Ca-alginate gel and uses lactic acid to form acetoin, a value-added product. The reaction is performed with a continuous, pH-dependent substrate feed under oxygenation. With titrated lactic acid yields of up to 91% and reaction times of ca. 6h was achieved. Using SPJ as titrant yields of 49% were obtained within 6h. In this deacidification process, with acetoin one value-added bio-based chemical is produced while simultaneously the remaining press juice can be used in applications that require a higher pH. Such, this system can be applied in a multi-product biorefinery concept to take full advantage of nutrient-rich SPJ, which is a widely available and easily storable renewable resource.

show abstract

Improvement of thermostable aldehyde dehydrogenase by directed evolution for application in Synthetic Cascade Biomanufacturing

Cited by 33 publications

References 42 publications

Cell‐free metabolic engineering: Biomanufacturing beyond the cell

Cell‐free metabolic engineering: Biomanufacturing beyond the cell

Modules for in vitro metabolic engineering: Pathway assembly for bio-based production of value-added chemicals

Deacidification of grass silage press juice by continuous production of acetoin from its lactate via an immobilized enzymatic reaction cascade

Contact Info

Product

Resources

About