Automated assay for screening the enzymatic release of reducing sugars from micronized biomass.

Microbial Cell Factories
David NavarroChristophe Bignon

Abstract

To reduce the production cost of bioethanol obtained from fermentation of the sugars provided by degradation of lignocellulosic biomass (i.e., second generation bioethanol), it is necessary to screen for new enzymes endowed with more efficient biomass degrading properties. This demands the set-up of high-throughput screening methods. Several methods have been devised all using microplates in the industrial SBS format. Although this size reduction and standardization has greatly improved the screening process, the published methods comprise one or more manual steps that seriously decrease throughput. Therefore, we worked to devise a screening method devoid of any manual steps. We describe a fully automated assay for measuring the amount of reducing sugars released by biomass-degrading enzymes from wheat-straw and spruce. The method comprises two independent and automated steps. The first step is the making of "substrate plates". It consists of filling 96-well microplates with slurry suspensions of micronized substrate which are then stored frozen until use. The second step is an enzymatic activity assay. After thawing, the substrate plates are supplemented by the robot with cell-wall degrading enzymes where necessary, and the wh...Continue Reading

References

Jun 13, 2000·Applied Biochemistry and Biotechnology·V S Chang, M T Holtzapple
Jun 13, 2002·Bioresource Technology·Ye Sun, Jiayang Cheng
May 2, 2003·Applied Biochemistry and Biotechnology·Stephen R DeckerMichael E Himmel
Oct 2, 2004·Biotechnology and Bioengineering·Zhizhuang XiaoAdrian Tsang
Dec 14, 2005·Biotechnology and Bioengineering·Alex BerlinJack Saddler
May 13, 2006·Biotechnology Advances·Y-H Percival ZhangJonathan R Mielenz
Aug 12, 2006·Biotechnology and Bioengineering·Shishir P S ChundawatBruce E Dale
Oct 13, 2006·Antonie van Leeuwenhoek·Antonius J A van MarisJack T Pronk
Feb 29, 2008·Biotechnology and Bioengineering·Shishir P S ChundawatBruce E Dale
Apr 1, 2008·The New Phytologist·Leonardo D GomezSimon J McQueen-Mason
May 15, 2008·Applied Biochemistry and Biotechnology·Yi ZhengBryan M Jenkins
Nov 15, 2008·Journal of Agricultural and Food Chemistry·Valerie Van CraeyveldChristophe M Courtin
Dec 25, 2008·Biotechnology for Biofuels·Isabelle Herpoël-GimbertMarcel Asther
Jun 9, 2009·Current Opinion in Biotechnology·Antoine MargeotFrédéric Monot

❮ Previous
Next ❯

Citations

May 7, 2013·Applied and Environmental Microbiology·Laetitia PoidevinSenta Heiss-Blanquet
Jul 10, 2012·Applied and Environmental Microbiology·Jean-Guy BerrinLaurence Lesage-Meessen
Dec 8, 2011·Microbial Cell Factories·Marie CouturierJean-Guy Berrin
Feb 10, 2012·Biotechnology for Biofuels·Scott J LeeSamuel P Hazen
Nov 26, 2013·Applied Microbiology and Biotechnology·Estelle BonninMarie-Christine Ralet
Jun 6, 2013·BioMed Research International·Panagiota M StathopoulouDimitris G Hatzinikolaou
Apr 4, 2014·Applied Microbiology and Biotechnology·Laetitia PoidevinEric Record
Feb 9, 2012·Bioresource Technology·Meng WangHuimin Zhao
Dec 23, 2015·Biotechnology for Biofuels·Marie CouturierMarie-Noëlle Rosso
Jul 26, 2012·Biotechnology Journal·Gernot Jäger, Jochen Büchs
Apr 14, 2016·Applied Biochemistry and Biotechnology·Amita SharmaSanjeev Kumar Soni
Oct 28, 2011·Bioresource Technology·Gabriela Ghizzi D SilvaXavier Rouau
Mar 31, 2012·Bioresource Technology·Holy RavalasonIsabelle Herpoël-Gimbert
Jan 8, 2017·Bioprocess and Biosystems Engineering·D TafouktJ-H Ferrasse
Dec 21, 2017·Scientific Reports·Brigitte ChabbertGabriel Paës
Jan 11, 2017·Scientific Reports·Ana VillaresBernard Cathala
Jan 12, 2017·Journal of Biological Engineering·Martina MühlmannJochen Büchs
Mar 30, 2019·Biotechnology for Biofuels·Camille Filiatrault-ChastelJean-Guy Berrin
Jan 13, 2016·The Journal of Biological Chemistry·Mickael LafondMarie-Line Garron
Jan 5, 2021·Environmental Microbiology·Nicolas ValetteMélanie Morel-Rouhier

❮ Previous
Next ❯

Software Mentioned

TRICEL

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.