Anaerobic detoxification of acetic acid in a thermophilic ethanologen

نویسندگان

  • A Joe Shaw
  • Bethany B Miller
  • Stephen R Rogers
  • William R Kenealy
  • Alex Meola
  • Ashwini Bhandiwad
  • W Ryan Sillers
  • Indraneel Shikhare
  • David A Hogsett
  • Christopher D Herring
چکیده

BACKGROUND The liberation of acetate from hemicellulose negatively impacts fermentations of cellulosic biomass, limiting the concentrations of substrate that can be effectively processed. Solvent-producing bacteria have the capacity to convert acetate to the less toxic product acetone, but to the best of our knowledge, this trait has not been transferred to an organism that produces ethanol at high yield. RESULTS We have engineered a five-step metabolic pathway to convert acetic acid to acetone in the thermophilic anaerobe Thermoanaerobacterium saccharolyticum. The first steps of the pathway, a reversible conversion of acetate to acetyl-CoA, are catalyzed by the native T. saccharolyticum enzymes acetate kinase and phosphotransacetylase. ack and pta normally divert 30% of catabolic carbon flux to acetic acid; however, their re-introduction in evolved ethanologen strains resulted in virtually no acetic acid production. Conversion between acetic acid and acetyl-CoA remained active, as evidenced by rapid (13)C label transfer from exogenous acetate to ethanol. Genomic re-sequencing of six independently evolved ethanologen strains showed convergent mutations in the hfs hydrogenase gene cluster, which when transferred to wildtype T. saccharolyticum conferred a low acid production phenotype. Thus, the mutated hfs genes effectively separate acetic acid production and consumption from central metabolism, despite their intersecting at the common intermediate acetyl-CoA. To drive acetic acid conversion to a less inhibitory product, the enzymes thiolase, acetoacetate:acetate CoA-transferase, and acetoacetate decarboxylase were assembled in T. saccharolyticum with genes from thermophilic donor organisms that do not natively produce acetone. The resultant strain converted acetic acid to acetone and ethanol while maintaining a metabolic yield of 0.50 g ethanol per gram carbohydrate. CONCLUSIONS Conversion of acetic acid to acetone results in improved ethanol productivity and titer and is an attractive low-cost solution to acetic acid inhibition.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Inactivation of Clostridium difficile in sewage sludge by anaerobic thermophilic digestion.

There has been an increase in community-associated Clostridium difficile infections with biosolids derived from wastewater treatment being identified as one potential source. The current study evaluated the efficacy of thermophilic digestion in decreasing levels of C. difficile ribotype 078 associated with sewage sludge. Five isolates of C. difficile 078 were introduced (final density of 5 log ...

متن کامل

Application of anaerobic digestion products of municipal solid food wastes in treating wastewaters

Anaerobic digestion is the breakdown of biodegradable organic material by microorganisms in the absence of oxygen or in an oxygen-starved environment.This technology is superior to the landfilling and also the aerobic composting. The aim of the present study was to examine whether the effluent Volatile Fatty Acids from the anaerobic acidogenesis of the food waste can be used du to its high valu...

متن کامل

An untapped bacterial cellulolytic community enriched from coastal marine sediment under anaerobic and thermophilic conditions.

A bacterial community with strong cellulose [filter paper (FP) and microcrystalline cellulose] degradation ability was isolated from the coastal marine environment. They were isolated under thermophilic (60 °C) and anaerobic cultivation conditions. The library of 16S rRNA gene clones revealed a total of 16 operational taxonomic units after 50 clones were surveyed. Sixty percent of the clones we...

متن کامل

Lignocellulose-derived thin stillage composition and efficient biological treatment with a high-rate hybrid anaerobic bioreactor system

BACKGROUND This study aims to chemically characterize thin stillage derived from lignocellulosic biomass distillation residues in terms of organic strength, nutrient, and mineral content. The feasibility of performing anaerobic digestion on these stillages at mesophilic (40 °C) and thermophilic (55 °C) temperatures to produce methane was demonstrated. The microbial communities involved were fur...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:

دوره 8  شماره 

صفحات  -

تاریخ انتشار 2015