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Process for production and quantitation of high yield of biobutanol

一种丁醇、微生物的技术，应用在定量评估丁醇,高效液相色谱领域，能够解决运行时间长、不能用于分析大量的样品等问题，达到减少时间的效果

Inactive Publication Date: 2010-12-29
RELIANCE LIFE SCI PVT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, even with these methods, the run time for the detection of butanol is very long (30-50 minutes)
Therefore, such methods cannot be used to analyze large numbers of samples

Method used

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  • Process for production and quantitation of high yield of biobutanol
  • Process for production and quantitation of high yield of biobutanol
  • Process for production and quantitation of high yield of biobutanol

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0117] Example 1: Biology and Growth Conditions

[0118] Clostridium acetobutylicum ATCC 10132 was grown in 125 ml anaerobic flasks containing 50 ml anaerobic sugar (AnS) medium with the following composition (gL -1 ): glucose (20.0), peptone (10.0), yeast extract (5.0), K 2 HPO 4 (3.0), NaCl(1.0), (NH 4 ) 2 SO 4 (1.0), CaCl 2 .2H 2 O(0.2), MgCl 2 .6H 2 O(0.2) and Na 2 CO 3 (1.0), pH 6.5.

[0119] The medium was sterilized (15 minutes, 121° C.) in glass vials sealed with butyl rubber stoppers. Fill the headspace with N 2 , and add Na 2 S.9H 2 O (0.02%) to remove traces of dissolved oxygen (Samuelov et al. 1991; Lee et al., 2000). The reduced medium was inoculated using a 2% inoculum size and incubated at 37 ± 1 °C for 96 h with intermittent gentle shaking.

Embodiment 2

[0120] Example 2: Method for evaluating butanol

[0121] After the desired incubation period, the culture was removed from the sealed vial using a sterile disposable syringe and centrifuged at 8000 xg for 10 minutes in an Eppendorf centrifuge (model 54151). Filter the supernatant through a 0.45 μ filter. At 37 °C, using acetonitrile and 0.5 mM H 2 SO 4 (1:9) as mobile phase and samples (20 μl) were analyzed by HPLC (Ehrlich et al, 1981) on a PRP 300X column (Hamilton) at a flow rate of 1.5 ml / min. Butanol was detected using an RI detector.

Embodiment 3

[0122] Embodiment 3: batch fermentation method

[0123] Various tested media for butanol production were: anaerobic sugar (AnS) medium (Isar et al., 2006); fortified Clostridium (RC) medium (Lin and Blaschek, 1983); soluble starch culture medium (SSM) (Moreira et al., 1981); alternative thioglycolic acid (AT) medium (Lin and Blaschek, 1983); potato starch medium (PSM) (Fouad et al., 1976). AnS was the best of the media tested, yielding 3.2 g / l of butanol (Table 1). Butanol production was optimized in AnS medium, where the influence of different physiological and nutritional parameters was investigated.

[0124] Table 1: Butanol production (g / l) in different media

[0125]

[0126] Subsequently, the effect of pH (5-7) and temperature (25-45°C) was studied in media selected for butanol production. The optimum pH for production was found to be 6.5 and 3.2 g / l of butanol was obtained within 84 hours (Figure 1). The results of the effect of different temperatures (25°C, 33°C...

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Abstract

The present invention provides a process for production of high yields of butanol by Clostridium acetobutylicum ATCC 10132. The process can be completed in a shorter span of time, using batch process through manipulation of various process parameters. The process can also be used for biomass based production of butanol. This paves the way for the strategic shift of major chemical industries from the hazardous routes of biofuel production to the biological, environmentally benign one.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of Indian Provisional Application No. 2544 / MUM / 2007, filed 24th December, 2007 and Indian Provisional Application No. 1367 / MUM / 2008, filed 30th June, 2008, which are incorporated by reference in their entirety Incorporated into this article. technical field [0003] The present invention relates to an improved process for the production of butanol in high yields using Clostridium acetobutylicum ATCC 10132. The present invention specifically reports strains with improved butanol tolerance under optimized conditions. The present invention also relates to a method for the rapid and efficient quantitative assessment of butanol during a fermentation process. In particular, the present invention relates to a high performance liquid chromatography (HPLC) method for the quantitative assessment of butanol produced by anaerobic fermentation using Clostridium acetobutylicum ATCC 10132. Backgr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/16
CPCC12P7/16Y02E50/10
Inventor 兰加斯瓦米·威贴伊萨尔·贾思明维尔玛·普拉迪普
Owner RELIANCE LIFE SCI PVT
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