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Shake Flask Fermentations

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Looking for additional gallons of ethanol fuel?

Laboratory instrument for fermentation validation

Shaken bioreactors are the most commonly used reaction vessels in biotechnology thanks to their simple design and handling. By using them together with our automated Gas Endeavour® III system, an online gas volume and flow measurement tool, you will be able to not only take advantage of the high experimental throughput, reduced costs and effort but also gain better insight and an increased understanding of the fermentation kinetics, thus allowing for optimisation of your bioethanol production process.

The anaerobic respiration rate is directly proportional to the metabolic activity of the fermenting microorganism and thus allows a continuous monitoring of the ethanol formation rate. Measuring respiration rates in shake flask cultures continuously enables you to see what is happening in real-time when it is actually happening. More information about the fermentation means more control options, which is the key to ensure a robust and high-yielding fermentation process. The Gas Endeavour provides you with a powerful laboratory tool to monitor shake flask fermentations, select the most suitable strains for production scale, evaluate yeast or bacteria activity, screen the additives and pre-treatment options, as well as develop an optimised operational procedure for fermentation processes.

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Instrument for strain screening and fermentation process optimisation

Areas of application

related to bioethanol production

Before the large-scale fermentation, fermentation process optimization is one of the most crucial parts to achieve a high yield production and a relatively low cost. Gas Endeavour III provides more information on the microbiological process in shake flask. With the help of the measurement curves, basic biological phenomena (e.g., microbial activity, microorganism respond to stress conditions, substrate limitation, product inhibition) can be reliably detected during process development.

Many strains of yeasts and bacteria are known to efficiently ferment sugars from various sources to ethanol. However, there is a constant quest for improving and selecting the best microorganisms for industrial application. Recent advances in strain development and improvement through adaptive laboratory evolution and/or metabolic engineering simplified and shortened the time to develop new strain variants in large numbers. Screening under relevant conditions is key for evaluating strain performance.

Apart from selecting a production strain, designing an appropriate fermentation medium and identifying the suitable operation conditions (e.g., agitation, pH temperature, inoculum size, incubation time) are of crucial importance to improve the efficiency of production for any fermentation process. Using shake flasks together with Gas Endeavour III allows a fast and an effective way to screen the important factors among several variables and also provide information on fermentation kinetics for process optimization.

In addition to sugars, several degradation products could be formed during the pre-treatment of lignocellulosic feedstocks that can negatively affect ethanol production. Characterise the toxic effects of inhibitory compounds (toxicity studies), assess the fermentability of lignocellulosic hydrolysates, as well as evaluate different detoxification strategies can be more easily performed with the help of Gas Endeavour III.

Statistically reliable results demand many parallel tests which considerably increase the amount of experimental effort required. Gas Endeavour III can be used to carry out 18 tests automatically during the same period in parallel and record it online in real time, thus significantly saving you time and money. Determination of yeast and bacteria activity, various technical approaches for process optimisation and enhancement of fermentation production can be made easy with Gas Endeavour III.

Benefits & advantages

Continuous and real-time measurements

The Gas Endeavour allows for continuous and on-line monitoring of alcoholic fermentation processes by continuously following the emission of carbon dioxide in real-time. Fully integrated and automatic, the Gas Endeavour significantly reduces labour and skill demands compared to competitive solutions. It produces high-quality data in real-time for extracting important kinetic information.

Direct and non-invasive integration with a sterile barrier

Universal fitting opportunities for convenient aseptic integration on new or existing equipment with a sterile barrier. Bottle nut and lid with twin connectors are available to easily connect to any Erlenmeyer flask with GL 45 thread in accordance with DIN 12380 / ISO 1773.

Automatic pressure and temperature compensation leading to high accuracy and reliability in analysis

Real-time temperature and pressure compensation minimise the impact of possible variations in measurement conditions and standardise data presentation satisfying the highest demands for data accuracy and precision.

Compact and modular design

The modular approach enables flexible system set-up, easy upgrading options and simple maintenance. The Gas Endeavour works as a stand-alone unit with 15 gas volume meters and can easily be further expanded by connecting multiple instruments with a network switch in order to satisfy demands and shorten development times.

Short set-up times and easy handling

Simple to use and easy to learn. The web-based software application makes setting up and monitoring experiments very easy. The Gas Endeavour also allows easy access from a remote location using any computer, smartphone or tablet.

Gas Endeavour® III

– a novel tool for process optimisation of bioethanol production

The Gas Endeavour III is a novel platform for analysing low gas volume and flow whenever there is a demand for high accuracy and precise measurements. The instrument can be used for research and industrial applications relating to: bioethanol fermentation, animal nutrition, wastewater, aerobic and anaerobic respiration, greenhouse gas emission, evaluation of microbial communities and more.

BPC Gas Endeavor MAX for both batch and continuous anaerobic and aerobic biodegradability testing
Reference

Scientific reference on bioethanol

Article title: “High bioethanol titre from Manihot glaziovii through fed-batch simultaneous saccharification and fermentation in Automatic Gas Potential Test System(AMPTS®)”

Authors: Anselm P. Moshi, Carla F. Creespo, Malik Badshah, Kenneth M.M. Hosea, Anthony Manoni Mshandete, Bo Mattiasson

Bioresource Technology (2014) 156, 348-356

Biotechnology, Department of Chemistry, Lund University (Sweden)

AMPTS® II was adapted to yeast ethanol fermentation and demonstrated to be an accurate, reliable and flexible device for studying the kinetics of yeast in simultaneous saccharification and fermentation (SSF) and fed-batch and simultaneous saccharification and fermentation (FB-SSF) of wild, non-edible cassava Manihot glaziovii. The correlation of CO2 derived ethanol agreed HPLC analysis (R2=0.99), hence recommended for fermentation kinetic studies.

Reference

Scientific reference on ethanol fermentation

Article title: “High temperature simultaneous saccharification and fermentation of starch from inedible wild cassava (Manihot glaziovii) to bioethanol using Caloramator boliviensis

Authors: Anselm P. Moshi, Ken M.M. Hosea, Emrode Elisante, G. Mamo, Bo Mattiasson

Bioresource Technology (2015) 180, 128-136

Biotechnology, Department of Chemistry, Lund University (Sweden)

The thermoanaerobe, Caloramator boliviensis was used to ferment starch hydrolysate from inedible wild cassava to ethanol at 60 oC. A raw starch degrading a-amylase was used to hydrolyse the cassava starch. During the fermentation, the organism released CO2 and H2 gases. An early prototype of Gas Endeavour® was used for monitoring and recording formation of these gaseous products. The bioethanol produced in stoichiometric amounts to CO2 was registered online in Gas Endeavour® and correlated strongly (R2=0.99) with values measured by HPLC.

Reference

Scientific reference on ethanol fermentation

Article title: “Production of bioethanol from unwashed-pretreated rapeseed straw at high solid loading”

Authors: Li Tan, Jia Zhong, Yan-Ling Jin, Zhao-Yong Sun, Yue-Qing Tang, Kenji Kida

Bioresource Technology (2020) 303, 122949

College of Architecture and Environment, Sichuan University; CAS Key Laboratory of Environmental and Applied Microbiology, Environmental Microbiology Key Laboratory of Sichuan Province, Chengdu Institute of Biology, Chinese Academy of Sciences (China)

AMPTS® II was used for ethanol fermentation using YPD medium with addition of acetic acid to study the inhibitory effect of acetic acid on the activity of yeast, S. cerevisiae KF-7. It appears to be essential to maintain acetic acid concentration below certain level for efficient ethanol fermentation.

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