FORM 2
THE PATENT ACT 1970
(39 of 1970)
&
The Patents Rules, 2003
\PROVISIONAL COMPLETE SPECIFICATION
(See section 10 and rule 13)
1. TITLE OF INVENTION PROCESS FOR THE PRODUCTION OF ETHANOL FROM
JATROPHA OILSEED CAKES FROM THE BIODIESEL PLANT.
2. APPLICANT(S)
(a) NAME: MR. B. CHANDRASEKHAR MR. MOHIT S. MISHRA
(b) NATIONALITY: INDIAN
(c)ADDRESS: 317, ABHYANKAR NAGAR, NAUPUR, 770010, MAHARASHTRA.
3. PREAMBLE TO THE DESCRIPTION

PROVISIONAL

COMPLETE

The tallowing specification describes the Invention.

The following spectfteafon particulariy describes Ihe invention and the raider in which it is to be performed.

4. DESCRIPTION {Description shall start from next page.)
5. CLAIMS (not applicable for provisional specification. Claims should start with the preamble — "I/we claim" on separate page)
6. DATE AND SIGNATURE (to be given at the end of last page of specification)
7. ABSTRACT OF THE INVENTION (to be given along with complete specification on separate page)
1 Nots: -
*Repeat boxes in case of more than one entry.
To be signed by the applicant!*) or by authorized registered patent agent , "Name of the applicant should be given in full, family name In the beginning . Con.olete address of the applicant should be given stating the postal index no,/code, *tate and country. 'Strike out the column which is/arajsot applicable
1499 MUM 2009
24 JUN 2009

TITLE- PROCESS FOR THE PRODUCTION OF ETHANOL FROM JATROPHA OILSEED CAKE FROM BIO-DIESEL PLANT
FIELD OF INVENTION
The present invention deals with a process for the production of ethanol from the oil cakes obtained from the biodiesel industry.
BACKGROUND AND PRIOR ART
The present invention describes a method for the process of producing ethanol from the byproduct waste cake of jatropha oilseeds obtained from the biodiesel production plant. Ethanol is a renewable substitute of petroleum fuels such as petrol and gasoline. Presently, major portion of the ethanol produced worldwide is produced by the fermentation of sugars obtained from molasses, cereals, and fruits. Much concern have arisen amongst the researchers of the world regarding the production of cellulosic ethanol form various cellulose-containing materials. Cellulose is an abundantly found carbohydrate and is found in many wastes such a sludge, paper and pulp, wood, textile waste etc. Utilization of cellulose, thus, for the production of ethanol may minimize the current dependence of the world on petroleum fuels and hfence making it less expensive.
Jatropha is a tropical plant and the oil obtained from its seeds is widely used for the production of biodiesel. The production of biodiesel from jatropha has been published in an article titled "Jatropha curcas L as a source for the production of biodiesel: A Cuban experience" by Jose Angel Sotolongol et al, wherein its has been mentioned that the waste press cake is produced as a byproduct of biodiesel production. The cake contains carbohydrates, fibres and rest of oil and is useful as organic fertilizer because the high content of nitrogen. Jatropha oil cakes can also be further processed and used to make biogas.
The present invention desribes a process for the production of ethanol from the jatropha oil cakes by the conversion of cellulose from thft cake into reducing sugars and the fermenting the sugars into ethanol so that it provides an efficient method to recycle and produce ethanol from the waste cellulose containing byproduct.
The process is useful for the Jatropha biodiesel industry since two important biofuels can be produced from the price of one raw material i.e. Jatropha oilseeds, by

setting up an integrated unit for the production of cthanol by fermentation in the biodiesel production plant.
Moreover, since the method for utilization of the oilseed cake for ethanol production has not been developed, as such the following invention is a novel method for the process. The process of obtaining ethanol from various cellulosic wastes have been worked out in the past, with reference to the works- "Systems and processes for cellulosic ethanol production" by Stephen L, Rush, and "Process for integrating cellulose and starch feed stocks in ethanol production'1 by Clifford Bradley. The present invention is novel by. its approach since it provides a specific and simpler method of cellulosic ethanol production from jatropha oilseed cakes
OBJECT OF THE INVENTION
The object of the invention is to provide a novel method of pretreatment and hydrolysis of the waste byproduct- oilseed cakes collected from the j atropha biodiesel industry, followed by the fermentation of the reducing sugars released in the hydrolyzed waste, to produce ethanol.

SUMMARY
The overall work deals with a novel and simplest method for the production of ethanol
from the oilseed waste cake, which lias been collected from the biodiesel industry. The
idea of using the waste byproduct from the industrial sector can be reutilized for
producing a novel method of ethanol production. This method basically describes the
recycle process of using renewable byproduct, which is easily produced inside the
industry.
The invention also describes the competitive comparison between the ethanol productions
from cellulose rich cotton waste already utilized and the oil cakes obtained as industrial
biodiesel waste.
This research describes the novel methods comprising of- conditions for pretreatment of
cellulosic wastes of oil cakes; temperature, incubation period, and acid concentrations of
the dilute acids for the hydrolysis of the pretreated cellulose containing oil cake waste;
the media components, incubation conditions for fermentation of the reducing sugars
released subsequently as a result of hydrolysis, to produce ethanol.
The pretreatment method includes adding 0.5 % H2SO4 to the biomass and autoclaving it
at 120°C for 1 hr. The hydrolysis was performed with 2% and 5% H2S04 at 55°C for 3
days and the estimation of rate of hydrolysis of the cellulose is done.
The fermentation method by Saccharomyces cerevisiae involves, preparation of media
broth for an anaerobic fermentation, temperature at 32 °C and an agitation at 110 rpm.
The estimation of ethanol showed a production of 8.5% ethanol v/v and 7.9% ethanol v/v
in jatropha oilseed cake broth and cotton waste broth respectively, and extraction of
ethanol is done by simple distillation method.

BREIF DESCRIPTION OF THE DRAWINGS
Figure No.l - Process of the ethanol production from the jatropha oilseed cake by hydrolysis and fermentation.
Figure No.2 — Comparison between amount of glucose released from the oil cake and cotton waste as a result of hydrolysis by 2% H2SO4 and 5% H2SO4.
Figure No. 3 - Comparison between the increase in the rate of hydrolysis with the increase in concentration of acid used for the hydrolysis.
Figure No. 4 - The day wise production of ethanol in the fermentation broth containing hydrolyzed oilseed cake waste and cotton waste.
Figure No. 5 - The day wise consumption of glucose in the fermentation broth.
DESCRIPTION
This invention is related to the process and methods applied to the production of ethanol from the waste cakes of jatropha oil seeds obtained after the extraction of oil in the biodiesel plant press.
One aspect of the invention is to provide a novel method of pretreatment of the waste in order to break the crystalline structure of the lignocellulose and remove the lignin to expose the cellulose and hemi cellulose molecules to facilitate the cellulose hydrolysis by either acids or enzymes. Depending on the biomass material, either physical or chemical pretreatment methods may be used. Physical methods may use high temperature and pressure, milling, radiation, or freezing—all of which require high-energy consumption. The chemical method uses a solvent to break apart and dissolve the crystalline structure. Due to the tough crystalline structure, the enzymes currently available require several days to achieve good results. Currently the cost of enzymes is also too high and research is continuing to bring down the cost of enzymes.

Keeping this view in mind, the present invention provides a method of steam presentment of the cellulosic oil cake waste in order to make the cellulose accessible for cither enzymatic or acid hydrolysis.
The temperature of the steam, the residence lime in the reactor, the size of the particles, moisture content and the concentration of the catalyst affect the steam pre-treatment. For rapid continuous processes, in order to allow adequate acid penetration, feedstock must also be reduced in size so that the maximum particle dimension is in the range of a few millimeters. Hence, the pretreatment method utilizes following simple steps- the mechanical reduction of the size of waste oil cake to finely ground powder; addition of 0.5% sulphuric acid to the ground cellulosic waste since a higher yield is obtained with a catalyst like H2SO4, and then autoclaving the mixture at a temperature of 120 °C and 15 psi pressure.
Hydrolysis is the process of breakdown of cellulose into cellobiose and glucose, which can be accomplished either by enzymes or by acid. Both enzymes and strong acids can do hydrolysis.
The enzymes are selective, results in a relatively high yield and the hydrolysis by enzymes is done at lower temperature. Most of the works describe the hydrolysis reaction of cellulose in wastes carried out with enzymes like cellulase and B-glucanase produced from a variety of microorganisms. However, for enzymes to work, they must obtain access to the molecules to be hydrolyzed. Due to the tough crystalline structure, the enzymes currently available require several days to achieve good results. Also, the high cost of the enzymes
Hydrolysis by acids involves concentrated and dilute acid hydrolysis, but the process of this invention utilizes dilute H2SO4 as the hydrolyzing agent due to its advantage over the concentrated acid process as it does not require deacidification after the hydrolysis no lower the pH and also its is more efficient than the. concentrated acid process. The present method of dilute acid hydrolysis is a novel approach to hydrolyze the cellulose and hemicellulose from the oilseed cakes.

The invention describes hydrolysis as a method of reacting the pretreated sample with 2%-5% H2SO4 at high temperature of 55°C for along duration of 3 days, known as "dilute acid hydrolysis" of cellulose. The hydrolysis is important because of the negligible concentration of reducing sugars in the unhydrolyzed waste, which is estimated to be 0.50 mg/ml, and can be increased to 18.60 mg/ml after applying the new approach of the acid hydrolysis.
The advantage of the new approach is a higher rate of hydrolysis and subsequent higher conversion rate of cellulose into reducing sugars with low energy consumption. The rate of hydrolysis of the pretreated oil seed cake waste is 0.238 mg/ml/hour when the hydrolysis is done with 2% H2SO4 which almost increases by 5% to 0.250 mg/ml/hour when the hydrolysis is done by 5% H2SO4. The efficiency of the new approach is also proved by the high cellulose conversion rate percentage of 68%, calculated as a result of the hydrolysis by 5% H2S04.
Another aspect of the present invention is the collection of the microbial strain (Saccharomyces cerevisiae) so as to use the strain for the process of overall fermentation after the preparation of the media containing the hydrolyzed waste sample. The strain was selected for its capacity produce ethanol to concentration as high as 18% of the broth by volume. It can use both simple sugar of glucose and disaccharide of sucrose. Saccharomyces cerevisiae is a generally recognized as safe (GRAS) organism and produces the necessary enzymes viz. zymase or invertase needed for ethanol production. The strain selected for the invention is collected from the brewing industry and stored under optimum conditions. The inoculum preparation for the fermentation is an important step. The strain selected must be incubated in sucrose or Saubourd's Agar Media at 37°C for at least 4 days. When the density of the yeast ceils in the liquid medium is adequate, the yeast may be used as the inoculum in the fermentation media. The size of the inoculum depends on the volume of the fermentation media and the amount of sugars to be fermented.

The invention also involves the batch fermentation of a media consisting of the pretreated and hydrolyzcd waste oil cake in order to convert the released sugars into ethanol. the conversion process being accomplished by the enzymes released by the Saccliaromyccs ecrevfsiae used as inoculums in the fermentation broth medium. Both the glucose released from the cellulose hydrolysis and other sugars released from hemicellulose hydrolysis are converted to ethanol by the same organism. In this method, the fermentation media may also involve the addition of salts and nutrients, to make the process more effective. The ideal conditions for the process are a temperature of 32°C, agitation of the broth at 110-rpm under anaerobic conditions.
The production of ethanol in the fermentation broth is estimated to be 8.5 % v/v at the end of 9 days of fermentation. It is found that, by this method at least 8 ml of ethanol can be produced from 30 gm of jatropha oilseed cake waste. The concentration of ethanol increases regularly in the broth along with a constant consumption of glucose by the yeast.
A comparison of the ethanol production from the oilseed cake with that from the cotton waste shows clearly that the ethanol production from the waste oil seed cake is more efficient with respect to several aspects including the hydrolysis rate, the cellulose conversion rate, and also the amount of glucose released at the end of the hydrolysis period, when the same pretreatment, hydrolysis and fermentation conditions are provided to each of them. Clearly, the method described in the invention is suitable more in the case of jatropha oilseed cake in contrast to the high cellulose containing cotton waste.
The possible reason behind this may be that the cellulose structure of the cotton is not easily accessible to the acid hydrolysis after the pretreatment process, but the same process when applied to the oilseed cake waste, a higher yield of the reducing sugars is achieved, because of the better hydrolysis rate.

It can be concluded that the present method of hydrolysis is more suitable for the oilseed cake used in the industrial research, even when it has lesser cellulose content than cotton.
The fermented media containing the ethanol is the product of the process, which is referred generally as beer. The separation and recovery of the ethanol is done by the conventional method of simple distillation, a process that leaves behind water and non¬volatile components of the beer. Any of the new distillation and ethanol recovery methods can be used in this invention.
The invention of this process is aimed at providing a technology that can be easily mtegrated in biodiesel industry, for the simultaneous production of ethanol from the byproduct oilseed cake it produces, using the novel and simpler methods described in the invention.