CN103397054A - Method for producing ethanol by enteromorpha fermentation - Google Patents

Method for producing ethanol by enteromorpha fermentation Download PDF

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Publication number
CN103397054A
CN103397054A CN2013103099122A CN201310309912A CN103397054A CN 103397054 A CN103397054 A CN 103397054A CN 2013103099122 A CN2013103099122 A CN 2013103099122A CN 201310309912 A CN201310309912 A CN 201310309912A CN 103397054 A CN103397054 A CN 103397054A
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enteromorpha
fermentation
ethanol
method disclosed
liquid
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韩玥君
宋建侠
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    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02EREDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
    • Y02E50/00Technologies for the production of fuel of non-fossil origin
    • Y02E50/10Biofuels, e.g. bio-diesel

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Abstract

The invention discloses a method for producing ethanol by enteromorpha fermentation. The enteromorpha, which is a common beach pollutant, is used as a substrate, and saccharomyces cerevisiae is used as a strain for the fermentation production of the ethanol; and compared with the traditional ethanol production processes using raw materials, such as sugarcane, corn and sweet potato, the method disclosed by the invention has the advantages that the raw material cost is reduced, and therefore the method is wide in generalizability. According to the method disclosed by the invention, the enteromorpha is subjected to enzymolysis in an acid condition, so that the crude fiber structure of the enteromorpha is destroyed to provide sufficient carbon resource for the fermentation process. According to the method disclosed by the invention, as the hydrolysis and the fermentation of the enteromorpha are continuously carried out, reducing sugar needed by fermentation can be supplemented and utilized in time, not only is the stagnation of the fermentation process caused by insufficient raw material prevented, but also the reduction of the fermentation efficiency caused by excessive raw material is avoided. According to the method disclosed by the invention, the fermentation condition of the enteromorpha is also optimized, and therefore the production process of the ethanol from the enteromorpha fermentation is efficiently achieved, the ethanol production by the enteromorpha fermentation is high in yield, and therefore another solution channel is provided for conquering energy crisis. The method disclosed by the invention has the characteristics of low cost, simple and convenient preparation, and wide application prospect.

Description

A kind of method of producing ethanol with fermenting enteromorpha
Technical field
The invention belongs to the bioenergy preparing technical field, be specifically related to a kind of method of producing ethanol with fermenting enteromorpha.
Background technology
As a kind of macro, Enteromorpha (Enteromorpha prolifera) is Chlorophyta (Chlorophyta) Ulvales (Ulvales) Ulvaceae (Ulvaceae) plant.In China, extensively distribute, especially at southeastern coast one band.In recent years, because the destruction of a large amount of burnings of the fossil oils such as oil and vegetation causes carbon dioxide content in atmosphere to increase, make a large amount of propagation of marine green algae such as Enteromorpha and cause the frequency of " green tide " and geographic range occurs to show a rising trend.Especially breaking out of 2008 Qingdao marine site Enteromorphas, a large amount of Enteromorphas is salvaged, and because high temperature rots to give birth to smelly, has had a strong impact on the urban environment in Qingdao.Meanwhile, China is the countries that petroleum resources are relatively deficient, the Imported oil just since 1993, and the external interdependency of energy demand constantly increases, and form is very urgent.China mainly utilizes the development of raw materials biomass resources such as stalk, waste wood, corn cob, and the exploitation of the marine alga energy is mainly produced to diesel oil as main take micro-algae.With respect to terrestrial plant, Enteromorpha belongs to unicellular lower eukaryote, but Enteromorpha has huge biomass, and breeding is being contained abundant bioenergy, so broad prospect of application is arranged rapidly.In order to realize the efficient utilization of Enteromorpha, nutritive ingredient to Enteromorpha is studied, and its each component content has been carried out to quantitative analysis, research shows that the nutritive ingredient of Enteromorpha is varied, and wherein a large amount of carbohydrate more makes Enteromorpha become industrial medium to become possibility.The objective of the invention is to process problem in order to solve Enteromorpha, realize simultaneously its efficient utilization.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of method of producing ethanol with fermenting enteromorpha, fast processing Enteromorpha on a large scale, and that the method has is with low cost, prepare characteristics easy, that have a extensive future.
The present invention produces the method for ethanol with fermenting enteromorpha.Comprise the steps:
1) with fresh water, rinse Enteromorpha, then carry out drying treatment; Dried Enteromorpha is pulverized, and sieved, be crushed into the waterside lichenin of 50 ~ 80 order sizes;
2) yeast saccharomyces cerevisiae is inoculated to activation in slant medium, 30 ℃, after cultivating 24 h, 200 r/min concussions are inoculated in sterilized liquid seed culture medium, and 30 ℃, the 200/min concussion is cultivated 24 h and is obtained the yeast saccharomyces cerevisiae seed liquor;
3) the Enteromorpha powder of pulverizing being added to water and be made into the suspension that concentration of substrate is 10 g/L, add the compound nutritional salt brine solution, is 4.0 ~ 5.5 with acid-alkali accommodation pH;
4) to step 3) add complex enzyme hydrolysis liquid in the suspension of gained, at 45 ~ 55 ℃ of lower enzymolysis 10 h, obtain hydrolyzed solutions;
5) 6 ~ 8:1 adds step 2 in hydrolyzed solution prepared by step 4) by volume) the yeast starter liquid for preparing, 28 ~ 35 ℃ of lower anaerobically fermentings 12 ~ 72 hours; Fermented liquid obtains bio-ethanol through distillation.
Above-mentioned steps 2) in, seed culture medium is the YPD slant medium: yeast extract 10 g, peptone 20 g, glucose 20 g, agar 15 g, water 1000 mL.
(the g/ml suspension) composed as follows of the compound nutritional salt brine solution in step 3): 0.15% urea, 0.1% sal epsom, 0.1% ammonium sulfate, 0.05% dipotassium hydrogen phosphate, 0.25% potassium primary phosphate.
Complex enzyme hydrolysis liquid cellulase and cellobiase in described step 4), the amount of its cellulase are 1.8 ~ 2.0g/100ml, and the add-on of cellobiase is 0.9 ~ 1.0g/100ml.
The present invention, take common beach pollutant Enteromorpha as substrate, produces ethanol take yeast saccharomyces cerevisiae as strain fermentation, with tradition, with raw material production ethanol such as sugarcane, corn, sweet potatoes, compare, and has reduced raw materials cost, has replicability widely.The present invention, by enzymolysis Enteromorpha under acidic conditions, destroys the coarse fiber structure of Enteromorpha, for fermenting process provides sufficient carbon source.In the present invention, the hydrolysis of Enteromorpha and fermentation are carried out continuously, make the required reducing sugar of fermentation in time to be supplemented and to utilize, and have both avoided insufficient raw material and make fermenting process stagnation, have avoided again too much reducing fermentation efficiency because of raw material.The present invention also is optimized the fermentation condition of Enteromorpha, has realized efficiently the process that fermenting enteromorpha is produced ethanol.Fermenting enteromorpha has produced the ethanol of high yield, for overcoming energy dilemma, provides another solution route.Therefore the present invention have with low cost, preparation is easy, the characteristics that have a extensive future.
Embodiment
Below in conjunction with embodiment, method of the present invention is described in detail.
Embodiment 1
1) with fresh water, rinse Enteromorpha, rinse the impurity such as salinity that its adhering on surface, silt, then carry out drying; The dried Enteromorpha of gained pulverizes and sieves, and is crushed into 50 order sizes;
2) get a ring yeast saccharomyces cerevisiae, inoculation activation in slant medium, after 24 h are cultivated in 30 ℃ of 200 r/min concussion, be inoculated in sterilized liquid seed culture medium (yeast extract 10 g, peptone 20 g, glucose 20 g, agar 15 g, water 1000 mL), 30 ℃ of 200 r/min concussion cultivated 24 h and obtained the yeast saccharomyces cerevisiae seed liquor;
3) the Enteromorpha powder that step 1) is obtained is put into fermentation unit, adds water and is made into the suspension that concentration of substrate is 10 g/L; To adding compound nutritional salt (by the amount of every liter of suspension) in fermentation system: the urea of 1.5g, the iron vitriol of 0.1g, the sal epsom of 1g, the ammonium sulfate of 1g, the dipotassium hydrogen phosphate of 0.5g, the potassium primary phosphate of 2.5g; With 2 M H 2SO 4Regulating pH is 4.0,45 ℃ of hierarchy of control stationary temperatures;
The present invention is optimized component and the concentration of composite salt, not only can regulate the buffer environment of suspension, and the arriving of hydrolysis result of ensuing prozyme improved; And above-mentioned composite salt can also make bacterial strain in follow-up fermenting process can bring into play maximum enzyme work.
4) add complex enzyme hydrolysis liquid: the amount of cellulase is 2.0g/100ml, and the add-on of cellobiase is 1.0g/100ml.First enzymolysis 10 h obtain hydrolyzed solution.
The applicant has carried out long-term research to kind and the proportioning of prozyme, has finally determined complex enzyme formula of the present invention, and by the effect of above-mentioned prozyme, in the time of can making the yeast fermentation hydrolyzed solution, the output of ethanol is improved significantly.
5) 6.0:1 adds the yeast starter liquid that has prepared in above-mentioned fermented liquid by volume, anaerobically fermenting 12 hours, each organizes ethanol content in supernatant liquor (ethanol, total reducing sugars, glucose, wood sugar, rhamnosyl) is 3.15%, fermented liquid obtains bio-ethanol through distillation, detects and shows that ethanol conversion is 0.151g/g (raw material).
Embodiment 2
1) with fresh water, rinse Enteromorpha, rinse the impurity such as salinity that its adhering on surface, silt, then carry out drying; The dried Enteromorpha of gained pulverizes and sieves, and is crushed into 50 order sizes;
2) get a ring yeast saccharomyces cerevisiae, inoculation activation in slant medium, after 24 h are cultivated in 30 ℃ of 200 r/min concussion, be inoculated in sterilized liquid seed culture medium (yeast extract 10 g, peptone 20 g, glucose 20 g, agar 15 g, water 1000 mL), 30 ℃ of 200 r/min concussion cultivated 24 h and obtained the yeast saccharomyces cerevisiae seed liquor;
3) the Enteromorpha powder that step 1) is obtained is put into fermentation unit, adds water and is made into the suspension that concentration of substrate is 10 g/L; To adding compound nutritional salt (by the amount of every liter of suspension) in fermentation system: the urea of 1.5g, the iron vitriol of 0.1g, the sal epsom of 1g, the ammonium sulfate of 1g, the dipotassium hydrogen phosphate of 0.5g, the potassium primary phosphate of 2.5g; With 2 M H 2SO 4Regulating pH is 5.0,50 ℃ of hierarchy of control stationary temperatures;
4) add complex enzyme hydrolysis liquid: the amount of cellulase is 2.0g/100ml, and the add-on of cellobiase is 1.0g/100ml.First enzymolysis 10 h obtain hydrolyzed solution.
5) 6.0:1 adds the yeast starter liquid that has prepared in above-mentioned fermented liquid by volume, anaerobically fermenting 48 hours.Each organizes ethanol content in supernatant liquor (ethanol, total reducing sugars, glucose, wood sugar, rhamnosyl) is 4.01%, and fermented liquid obtains bio-ethanol through distillation.Above process implementation ethanol conversion be 0.232g/g (raw material)
Embodiment 3
1) with fresh water, rinse Enteromorpha, rinse the impurity such as salinity that its adhering on surface, silt, then carry out drying; The dried Enteromorpha of gained pulverizes and sieves, and is crushed into 70 order sizes;
2) get a ring yeast saccharomyces cerevisiae, inoculation activation in slant medium, after 24 h are cultivated in 30 ℃ of 200 r/min concussion, be inoculated in sterilized liquid seed culture medium (yeast extract 10 g, peptone 20 g, glucose 20 g, agar 15 g, water 1000 mL), 30 ℃ of 200 r/min concussion cultivated 24 h and obtained the yeast saccharomyces cerevisiae seed liquor;
3) the Enteromorpha powder that step 1) is obtained is put into fermentation unit, adds water and is made into the suspension that concentration of substrate is 10 g/L; To adding compound nutritional salt (by the amount of every liter of suspension) in fermentation system: the urea of 1.5g, the iron vitriol of 0.1g, the sal epsom of 1g, the ammonium sulfate of 1g, the dipotassium hydrogen phosphate of 0.5g, the potassium primary phosphate of 2.5g; With 2 M H 2SO 4Regulating pH is 5.5,55 ℃ of hierarchy of control stationary temperatures;
4) add complex enzyme hydrolysis liquid: the amount of cellulase is 2.0g/100ml, and the add-on of cellobiase is 1.0g/100ml.First enzymolysis 10 h obtain hydrolyzed solution.
5) 7.0:1 adds the yeast starter liquid that has prepared in above-mentioned fermented liquid by volume, anaerobically fermenting 72 hours.Each organizes ethanol content in supernatant liquor (ethanol, total reducing sugars, glucose, wood sugar, rhamnosyl) is 3.33%, and fermented liquid obtains bio-ethanol through distillation.Above process implementation ethanol conversion be 0.203g/g (raw material)
Embodiment 4
1) with fresh water, rinse Enteromorpha, rinse the impurity such as salinity that its adhering on surface, silt, then carry out drying; The dried Enteromorpha of gained pulverizes and sieves, and is crushed into 80 order sizes;
3) get a ring yeast saccharomyces cerevisiae, inoculation activation in slant medium, after 24 h are cultivated in 30 ℃ of 200 r/min concussion, be inoculated in sterilized liquid seed culture medium (yeast extract 10 g, peptone 20 g, glucose 20 g, agar 15 g, water 1000 mL), 30 ℃ of 200 r/min concussion cultivated 24 h and obtained the yeast saccharomyces cerevisiae seed liquor;
4) the Enteromorpha powder that step 1) is obtained is put into fermentation unit, adds water and is made into the suspension that concentration of substrate is 10 g/L; To adding compound nutritional salt (by the amount of every liter of suspension) in fermentation system: the urea of 1.5g, the iron vitriol of 0.1g, the sal epsom of 1g, the ammonium sulfate of 1g, the dipotassium hydrogen phosphate of 0.5g, the potassium primary phosphate of 2.5g; With 2 M H 2SO 4Regulating pH is 5.0,60 ℃ of hierarchy of control stationary temperatures;
4) add complex enzyme hydrolysis liquid: the amount of cellulase is 2.0g/100ml, and the add-on of cellobiase is 1.0g/100ml.First enzymolysis 10 h obtain hydrolyzed solution.
5) 6.0:1 adds the yeast starter liquid that has prepared in above-mentioned fermented liquid by volume, anaerobically fermenting 72 hours.Each organizes ethanol content in supernatant liquor (ethanol, total reducing sugars, glucose, wood sugar, rhamnosyl) is 2.78%, and fermented liquid obtains bio-ethanol through distillation.Above process implementation ethanol conversion be 0.134g/g (raw material).

Claims (4)

1. a method of producing ethanol with fermenting enteromorpha, comprise the steps:
1) with fresh water, rinse Enteromorpha, then carry out drying treatment; Dried Enteromorpha is pulverized, and sieved, be crushed into the waterside lichenin of 50 ~ 80 order sizes;
2) yeast saccharomyces cerevisiae is inoculated to activation in slant medium, 30 ℃, after cultivating 24 h, 200 r/min concussions are inoculated in sterilized liquid seed culture medium, and 30 ℃, the 200/min concussion is cultivated 24 h and is obtained the yeast saccharomyces cerevisiae seed liquor;
3) the Enteromorpha powder of pulverizing being added to water and be made into the suspension that concentration of substrate is 10 g/L, add the compound nutritional salt brine solution, is 4.0 ~ 5.5 with acid-alkali accommodation pH;
4) to step 3) add complex enzyme hydrolysis liquid in the suspension of gained, at 45 ~ 55 ℃ of lower enzymolysis 10 h, obtain hydrolyzed solutions;
5) 6 ~ 8:1 adds step 2 in hydrolyzed solution prepared by step 4) by volume) the yeast starter liquid for preparing, 28 ~ 35 ℃ of lower anaerobically fermentings 12 ~ 72 hours; Fermented liquid obtains bio-ethanol through distillation.
2. the method for claim 1, is characterized in that described step 2) in seed culture medium be the YPD slant medium.
3. the method for claim 1, is characterized in that compound nutritional salt brine solution composed as follows in described step 3): 0.15% urea, 0.1% sal epsom, 0.1% ammonium sulfate, 0.05% dipotassium hydrogen phosphate, 0.25% potassium primary phosphate.
4. the method for claim 1, is characterized in that in the step 4) of described step, complex enzyme hydrolysis liquid is the aqueous solution of cellulase and cellobiase, and the amount of its cellulase is 1.8 ~ 2.0g/100ml, and the amount of cellobiase is 0.9 ~ 1.0g/100ml.
CN2013103099122A 2013-07-22 2013-07-22 Method for producing ethanol by enteromorpha fermentation Pending CN103397054A (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103695333A (en) * 2013-11-21 2014-04-02 上海海洋大学 Stenotrophomonas FQ1 strain and its screening method and use
CN103695334A (en) * 2013-11-21 2014-04-02 上海海洋大学 Bacillus FH3 strain and its screening method and use
CN103710394A (en) * 2013-12-10 2014-04-09 南昌大学 Method for producing bioethanol by utilizing alga residues obtained after oil extraction

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101638671A (en) * 2009-08-27 2010-02-03 烟台海岸带可持续发展研究所 Method for preparing bioethanol by using enteromorpha as raw material
CN101864455A (en) * 2010-04-11 2010-10-20 中国海洋大学 A kind of oxidative degradation pretreatment enteromorpha as raw material that utilizes carries out the method that alcohol fuel transforms
CN102100398A (en) * 2011-01-25 2011-06-22 中国烟草总公司郑州烟草研究院 Preparation method of enteromorpha ethanol extract and application thereof in cigarette
CN102234663A (en) * 2010-05-05 2011-11-09 上海海洋大学 Method for preparing fuel ethanol

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101638671A (en) * 2009-08-27 2010-02-03 烟台海岸带可持续发展研究所 Method for preparing bioethanol by using enteromorpha as raw material
CN101864455A (en) * 2010-04-11 2010-10-20 中国海洋大学 A kind of oxidative degradation pretreatment enteromorpha as raw material that utilizes carries out the method that alcohol fuel transforms
CN102234663A (en) * 2010-05-05 2011-11-09 上海海洋大学 Method for preparing fuel ethanol
CN102100398A (en) * 2011-01-25 2011-06-22 中国烟草总公司郑州烟草研究院 Preparation method of enteromorpha ethanol extract and application thereof in cigarette

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103695333A (en) * 2013-11-21 2014-04-02 上海海洋大学 Stenotrophomonas FQ1 strain and its screening method and use
CN103695334A (en) * 2013-11-21 2014-04-02 上海海洋大学 Bacillus FH3 strain and its screening method and use
CN103695334B (en) * 2013-11-21 2015-08-26 上海海洋大学 A kind of genus bacillus FH3 bacterial strain and application
CN103695333B (en) * 2013-11-21 2015-08-26 上海海洋大学 A kind of Stenotrophomonas FQ1 bacterial strain and application
CN103710394A (en) * 2013-12-10 2014-04-09 南昌大学 Method for producing bioethanol by utilizing alga residues obtained after oil extraction
CN103710394B (en) * 2013-12-10 2016-05-11 南昌大学 A kind of method of utilizing oil-extracted algae slag to produce bio-ethanol

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Application publication date: 20131120