Propuesta Investigación IFS Ácido Cítrico
Propuesta Investigación IFS Ácido Cítrico
Propuesta Investigación IFS Ácido Cítrico
IFS also makes available other kinds of support to grantees in order that the young researchers may develop new
skills, realise research goals, and ultimately become an established member of the international scientific community.
This kind of support includes providing professional contacts for the grantee, offering research support in the form of
literature searches and access to databases, and organising IFS workshops and regional meetings. Supplementary
travel grants may be awarded enabling the grantee to attend scientific meetings. An important service provided by IFS
is the purchase and delivery of equipment and supplies to grantees. Further information about the supporting services
mentioned here will be made available to researchers with approved project proposals.
IFS grants are awarded on an individual basis. However, as the importance of teamwork is recognised by IFS,
separate applications from different members of the same research team will be considered.
1
Not qualifying for support are countries in Eastern and South Eastern Europe (including Turkey and Cyprus) or the former Soviet Union.
2
Chinese applicants have to be younger than 30.
3
Applicants are encouraged to send a copy of their application to the IFS Member Organisation (MO) in the applicant's home country.
Contact information for MOs is available on the IFS Homepage: www.ifs.se.
3
A multitude of disciplines and topics are supported by IFS. However, all applications must be related to the biological
resource base. The graphic below is an indication of the diversity of research topics that IFS supports, it is by no
means all-inclusive:
GRANT AGREEMENT
When an application is approved, a formal agreement is made between IFS, the grantee, and the Institution (or similar
authority) of the grantee. The grant can be transferred partly or entirely to the grantee's Institution, or it can be kept in
Stockholm for purchases to be made by IFS on behalf of the grantee. The grantee’s Institution undertakes to
administer the grant and guarantee that there exists the laboratory space, salaries, and other facilities necessary for
the project. Upon completion of the project, instruments and other equipment (with the exception of computers)
purchased with the grant money will normally become the property of the Institution. Whilst throughout the application
process, the monetary instrument is the United States dollar, the grant is ultimately administered in Swedish Crowns
and so, as a result of currency fluctuations, the actual amount allocated to the grantee’s project may be slightly more
or less than the US dollar amount.
At the end of the project period, the grantee must send a research report and a report of project expenditures.
While IFS does not claim rights to any publications, inventions, or patents arising out of a project, the grantee must
keep IFS informed of any possible applications of the research results and acknowledge IFS in publications resulting
from the research. Copies of publications must be submitted to IFS. Confidentiality will always be respected by IFS
and its collaborating international experts.
GRANT RENEWAL
An IFS grant may be renewed twice. A renewal application must be submitted together with a research progress
report and the report of project expenditures and is subject to the evaluation of the Scientific Advisers before approval.
Guidelines for reports and renewal grant application forms are available from IFS. No more than a total of three grants
will be awarded to a single researcher.
AWARDS
Grantees whose work is of high quality will be eligible for one of several annual awards. In addition, every three years
the Sven Brohult Award is given to a (former) grantee whose research effort has been outstanding.
1 APPLICANT
Your title click on the arrow in the drop-down list, and choose one option
Sex click on the arrow in the drop-down list, and choose one option
Your name as it would State your name as it would appear in a scientific publication. If you have several given
appear on your names, write out only one and use initials for the others.
publications:
Date of birth Please use numbers for months (ie 1 through 12), not text.
Most recent degree Provide the title of your most recent and most advanced academic degree. More details
regarding this and other degrees is asked for in section 6.1.
2 RESEARCH PROJECT
Title of your project: Keep the title of the project short and simple. The title should clearly describe the basic
objectives of the proposed project and be understandable by a scientist who is not an expert
in your field.
Short summary of the Give a short summary (maximum 250 words) of the proposed project, sufficiently informative
project: for other scientists who are not necessarily in your field of expertise. The summary should
contain an overview of the project, followed by the objectives and a brief description of how
these will be accomplished, with expected outputs. Considerable attention should be given to
the preparation of this item. It is suggested that you write this item last. Include an estimated
budget in the space below your summary (this should be the same as in section 11).
3 SIGNATURES
Both you and the Head of your Institution are required to sign the application form before a decision can be made by IFS.
You must print a copy of the first page of the application form after you have filled in the above items (1 & 2) and then both
you and the Head of your Institution must sign it. The first page of the application must be stamped with the
Institution's official stamp and then sent to IFS by ordinary mail. When you have completed the form, send the whole
document back to IFS as an attachment in an e-mail (Use the e-mail address: [email protected]).
4 CONTACT INFORMATION
Under the three headings Comprehend, Speak and Write, check the box that most correctly describes your skill in each of
the named languages.
6 YOUR EDUCATION
Give details on your most recent scientific education. Besides the names and addresses of the Institutions attended,
also give the dates of the courses, the names of your supervisors, and full information about the academic degrees you
obtained during these periods. Also give information about any fellowships you have received.
6.1 Major scientific education and academic degrees
Month/Year:
Please use the following format: MM/YY. For example, August 2002 would be entered as 08/02.
Names and addresses of educational establishments:
Please give the full postal address of the Institution. Write the address in the order: Department, Faculty or
Institution, University, PO Box or street address, city, country.
Degrees:
Here you should choose the degree you received at this educational establishment. If the degree you received
is not listed here, but is equivalent to one listed as an English, French or Spanish degree, choose that. If you
do not know what it is equivalent to in English, French or Spanish degrees, use section 6.2.
Specialisation: Indicate the area of specialisation for this degree.
Fellowship / Study grant from:
Give information about any fellowships or study grants you received to do this degree.
7 EMPLOYMENT
8.2 Describe your results and experience related to the proposed research project.
Provide information about results and experience you have obtained which is relevant to the proposed research
project.
8.3 Describe ongoing work at your Institution related to the research project.
Provide information about the scientific work at your Institution in the area of the proposed research project; also
mention any present or planned co-operative research with other Institutions.
9.1 Provide a background and justification for your proposed research. In addition to identifying the problem to be
addressed by the research, this section should also identify the specific stakeholders in this problem.
9.2 Provide a short and up-to-date summary of the present status of scholarly knowledge relevant to the research
you propose. Make reference to local and world-wide literature.
9.3 Provide the references which support the information in 9.2 (author, title, journal/book, volume, year, pages).
9.4 State the specific (short-term) objective(s) of the project and how they relate to the general (long-term)
objectives and the questions that the project will attempt to answer.
9.5 State the scientific hypothesis to be tested. This is a statement of research objectives in a way which can be
tested by experimentation.
9.7 Applicants for IFS research support are asked to justify the relevance of their research in terms of local and
national socio-economic and environmental conditions. IFS encourages applicants to engage in a dialogue
with potential end-users and other stakeholders to define research objectives that meet these criteria when
designing the research proposal. In this section state and justify the relevance of the project to the country or
region. Indicate the relevance in relation to national priorities or in a regional context and mention, if any,
economic, social or environmental benefits or consequences of the research project. You may also mention
potential commercial applications of your research.
9.8 Applicants with projects that produce data for statistical analysis are strongly advised to contact a
statistician/biometrician during the planning stage of the research project in order to make sure that the design the
project permits appropriate statistical analysis and interpretation of the data that will be collected.
9.9 Provide details on your experimental design or plans for trials and observations, indicating plot layout and sizes
(or equivalent information), treatments, replications and statistical method(s) to be used to analyse the data.
9.10 Describe the statistical methods that you will use to analyse your data. Provide the names of any software
programmes that you will be using.
9.11 The research plan is an extremely important part of the proposal, and IFS Scientific Advisers attach great
importance to it. It should be directed to specialists in the field. It should cover a research period of 1-3 years
(the budget - item 11 - has to correspond to this period). Do not repeat the summary you have given in item 9.2
or the statements you have already made in 9.4 or 9.6. Describe here (do not simply make a list of activities)
what will be done (observation work, laboratory experiments, participant observation, surveys etc) and how it
will be done, including the following points:
Methods and activities to be carried out to solve the problems and answer the questions stated in 9.4-9.5.
Which parameters will be studied and which measurements will be taken; which data will be collected and
how – do not simply make reference to a publication to substitute for a methodology description unless you
are referring to a basic methodology which is well-known to non-specialists in your field as well as
specialists.
Criteria for choosing sites and samples, sampling methods and sample sizes, Latin names of species. Provide
sources of germplasm where appropriate. If you will use a questionnaire in your research, you are
required to send a draft copy of your questionnaire to IFS with the completed application form.
Describe your field trial design (if appropriate). In some cases you may find it necessary to send a diagram to
IFS as an attachment. If you do, please refer to that diagram in this space.
Following the completion of an IFS supported research project, grantees are encouraged to make their
research results available in fora that encourage local organisations, individuals, governmental bodies, etc to
understand and use the benefits of the grantees’ research. Applicants are strongly encouraged to include a brief
plan in this section that indicates how their results will be made available to local stakeholders (eg farmers, local
industry, policy makers, natural resource users). If applicants wish to organise a small workshop/seminar, the
publication of a pamphlet, etc for these purposes, a budget item should be included in section 11.
IMPORTANT! If you are working with or producing genetically-engineered organisms, dangerous products, or are
carrying out experiments that may raise ethical issues, you should mention if and how safety regulations and
international standards will be taken into consideration. If you will solicit information from human subjects (using
interviews, questionnaires, etc), explain how you will protect the confidentiality of those individuals.
10.1 List here equipment and facilities, etc, necessary for your project which will be provided by the Institution where
the work will be carried out. This is important for a proper evaluation of the application. Spare parts or accessories
for these apparatus may be requested in the budget if they are necessary. The use of equipment included in this
list must be agreed upon by the Head of Institution; this is implicit in his/her signature in 3.1.
10.2 List any other funds (national or international) that you or your Institution have obtained or applied for in the area
of the proposed research project. State funding organisation, amounts, date of approval, and length of funding
period.
11 BUDGET
The total budget (items 11.1 - 11.6) should not exceed USD 12,000. The budget items must be relevant to the proposed
research plan and should be justified under item 11.6. Vehicles cannot be purchased from the IFS grant. IFS DOES NOT
FUND INSTITUTIONAL OVERHEADS OR HONORARIA.
11.1 List each item of equipment which you request. Equipment is considered to be items that have an individual cost
of USD 500 or more and a life duration of more than 3 years. Specify and describe each item, and give to the
best of your knowledge: manufacturer, model/type, accessories if needed. Give a realistic estimate of the cost for
each item. Include freight charges, insurance, and taxes for items to be imported. Consult, if possible, a senior
technician for advice on the most appropriate equipment for your project.
Please remember that IFS must follow all international trade agreements and trade embargos. In practice, this
means that IFS does not purchase and transfer equipment or materials between countries when such transfers
are restricted by one or both countries.
11.2 List categories and significant items of expendable supplies (glassware, solvents, feed, reagents, etc) and give a
cost estimate for each category and significant items. Include freight charges, insurance and taxes for items to be
imported.
11.3 List categories of literature (eg books, reprints, and photocopies) or documentation, and give a cost estimate for
each category. Recommended are books rather than subscriptions to journals since subscriptions will expire as
soon as IFS support ends. However, abstract reviews or membership in a scientific society publishing a journal
may be considered (please note that the grantee is expected to organise the purchase of items listed under 11.3,
not the IFS purchasing department). Literature searches may be included under item 11.3. IFS does not provide
funds to cover costs associated with publishing grantees' own scientific work.
11.4 Only local travel costs (for sample collection, field visits, participant observation, surveys etc) may be requested
under this item. Give destination and means of travel. The need for local travel has to be explained in 11.6 and a
specific calculation of travel costs must be included. If sites are far away from your institute, their choice has to be
justified under item 11.6. Travel costs to conferences, scientific meetings, etc, should not be included.
11.5 Extra manpower costs must be justified and their costs specified under item 11.6. They should only include
manual labour costs which are necessary for the execution of the project, eg, labourers for field work, animal
care, tedious preparations, translators, guides etc. The applicant cannot be subsidised with an honorarium, salary,
etc.
11.6 List the items you request funding for in the budget in US DOLLARS (11.1-11.5). Describe their purpose and
function and justify their need in relation to the activities mentioned in the research plan. Give explanatory details
for amounts requested under 11.4 and 11.5., and indicate governmental/official rates for 11.4 and 11.5.
Unexplained or insufficiently explained items may be cause for failure of the application or reduction of the grant
award.
Please state how you learned about IFS and from where you accessed the application form.
Short summary of project: Tropical zone of Cochabamba, Bolivia, is being subject to eradication of coca plantations.
Peasant's economy has already been severely affected. Alternative agricultural efforts, conducted with international
cooperation assistance, have reached only limited economic results. Banana production has been greatly enhanced, but
exports are less than 10% of present availability. Surplus and discarded banana prices make banana pulp industrialisation
attractive. Banana producer peasant associations are highly interested to find new markets. Citric acid production from
banana pulp sugars is presented as an alternative to add value to available raw materials and to reduce actual Bolivian
imports. Most world citric acid production is carried out by batch - submerged fermentation of molasses with Aspergillum
niger. It is proposed to initiate Bolivian experience in this field by screening Aspergillus niger citric acid producing strains
obtained from known collections on productivity in banana liquefied pulp substrates by surface fermentation experiments
and to optimise citric acid production of most promising one by studying best initial substrate composition and best
fermentor operating conditions during submerged batch fermentation. Effect of Sugars concentration, Nitrogen, Magnesium,
Phosphor, Copper, Iron and Manganese will be investigated, together with temperature, initial pH, aeration and stirring.
3. SIGNATURES You cannot sign this page on your computer. Print this first page. The Applicant and the Head of
Institution must sign it. It should be stamped by the institution and sent (THIS PAGE ONLY) by
airmail to IFS. When the form is completed, email the entire form to IFS at [email protected].
Name, title and official position: Manuel Olave Sarmiento, Dr., Rector
(please type)
Country: Bolivia
Telephone numbers at the Institution: Your direct no: 591 4 426 8287
(include area code but not country code) Institution’s general no: 591 4 426 8287
Fax no: 591 4 426 8288
Your e-mail address at this Institution: [email protected]
4.2 If you will be carrying out your research at an Institution or location different from the one provided in 4.1,
please provide the full address for your research location in the space below:
Full address:
(Include Department, Faculty
or Institution, University, PO
Box or street address, and
City, if relevant)
Country:
Is this your preferred mailing address? Yes No
Telephone numbers at this address: Your direct no:
(include area code but not country code)
Institution’s general no:
Fax no:
Your e-mail address at this place:
The purpose of your stay here:
Until what date is this address valid:
Further remarks:
Not easily
Not easily
Easily
Easily
Easily
English:
French:
Other:
Other:
Specialisation:
Name of your Fellowship /
Superviser: study grant from:
Employer:
Position:
8.2 Describe your results and experience related to the proposed research project
8.4 Research partners and/or relevant contacts already established by you; give names, full addresses, e-mail, field
of specialisation, and (if applicable) the specific role of partner researchers in this IFS project
8.5 Additional contacts you want to establish; give names, full addresses and field of specialisation
9.2 Give an up-to-date summary of scholarly knowledge in the field of the project
Citric acid is mainly produced by submerged fermentation of molasses (beet and sugarcane) with A. niger(1). The
fermentation process has been introduced industrially in 1919 in Belgium. Submerged fermentation has replaced superficial
fermentation. Highly productive A. niger strains have been developed, with yields over 90%, with regard to sugar
consumption, in 4 to 10 days(2). The process has been optimized in many laboratories(3,4). Although detailed metabolic
mechanisms are not completely known, the biphasic fermentation process is relatively well understood(5,6). Several
carbohydrate-containing substrates have been proposed(7,8,9). Only one reference has been found concerning banana
extract(10). Successful citric acid production depends mainly on the A. niger strain employed, proper oxygen transfer, that
depends on the combination of medium hydrodynamic properties, aeration, stirring and fermentor geometry(11) and proper
substrate formulation (carbohydrate contents, added nitrogen containing compounds and metallic salts and eventually
removal of inhibiting trace metallic cations)(12,13,14). Morphology of growing A. niger under suitable conditions for citric
acid production is well known(1, 14,15,16).
9.7 State the relevance of the project in relation to local and regional environmental and socio-economic
conditions
Coca plantations eradication in the tropical zone of Cochabamba has already caused economic disequilibria among
peasants. Alternative crops promoted by international agencies have reached limited impacts. Banana production has been
greatly enhanced with export varieties. Less than 10% of actual production is exported, and generated surpluses have
depleted prices. Among several adding value alternatives, citric acid production from banana pulp appears as an
economically attractive alternative, although at a small national level. Bolivian imports of citric acid amount 500 MT/year at
0.5 Million $US/year. Local citric acid production could replace actual imports.
Peasant banana producers are associated and are looking for new markets for their production.
An experimentally supported feasibility study for the production of banana based citric acid will be a benefit to them and the
state institutions that are pursuing new economic alternatives for the zone.
9.8 Research design and data analysis
Grantees with projects that primarily produce data for statistical analysis Have you contacted Yes
are strongly advised to contact an expert in statistical analysis during the an expert in
planning stage of the research project in order to make sure that the statistical analysis? No
design of the project allows for appropriate analysis. Not applicable
If Yes, Name and Institution:
9.9 Give reference to relevant literature from which you take your research methodology, statistical methods and
research or experimental design
Sikander Ali et al, Production of citric acid by Aspergillus Niger using cane molasses in a stirred fermentor, Electronic
Journal of Biotechnology, ISSN 0717-3458, Vol. 5, Nº 3, December 2002, p. 258-271
M. Moresi, E. Parente, Production of Organic Acids, Academic Press, 1999
9.10 Which methods will you use to analyse the information that you gather? Refer also to any software you will be
using.
Average of three (productivity screening) or two (submerged fermentation) independent experimental determinations under
the same conditions.
10 carefully selected mutant Aspergillus niger strains will be purchased from known sample collection holders,
that will be sub cultured monthly and maintained in potato dextrose agar at 4 ºC.
Conidia solutions of 4-7-day-old cultures from potato dextrose slants incubated at 30 °C will be used as
inoculants. The initial cell concentration will be controlled by spectrometry (5*10^7 conidia/ml).
Surface fermentation will be carried out with 100 ml liquefied banana pulp containing 15% total sugars during 7-
10 days at 30 ºC in 250 ml Erlenmeyer flasks held in a wrist action shaker, six at a time. 10 ml liquid samples
will be taken daily and analysed for remaining sugars and citric acid formed. Micelial growth will be also
qualitatively monitored.
By comparison of reported values of banana pulp composition and pre-treated molasses composition for citric
acid fermentation, at 15% ww sugar content, banana pulp is lower in Nitrogen content, higher in Fe, K, Mn and
has approximately the same amounts in Mg, Zn, Ca, Cu and P, among the most important chemical species
studied in this fermentation process.
Too high Fe and Mn values could be neutralized by K4Fe(CN)6 or by cation exchange. Too low Nitrogen
content could be corrected by urea or NH4NO3.
Eventually, liquefied banana pulp will be chemically adjusted prior to surface fermentations, to approximate pre-
treated molasses composition.
Liquefied banana pulp will be prepared by blending Cavendish banana edible part with distilled water to 15%
total sugar content. The liquid will be filtered, sterilised and stored at 4 ºC. 3 litres will be prepared each month
for all needed experiments.
Sugars will be determined as total carbohydrates by the phenol sulphuric acid method. Citric acid will be
determined by titration with NaOH according to the AOAC method. Periodically, the presence of citric acid will
be verified and quantified by the pyridine acetic anhydride method.
The ten strains will be tested three times. Required time: 2-3 months.
Afterwards, best temperature, initial pH, initial sugars composition and stirring/aeration conditions will be found.
All experiments will be realised twice. About eighty different runs will be performed during 7 to 10 days each.
Sugars content and citric acid will be analysed daily by the above-described methods. Micelial mass growth will
be also measured daily by centrifugal separation, drying and weighing of fermenting medium samples.
10.2 List any other funds that you or your Institution have obtained or applied for this project
Donor Time Frame Amount (USD)
0
0
0
If you have co-funding for your research, please give details of why IFS support is needed to complement your other support:
Have you at any time applied for or received support from TWAS or IFS for this research? If so, give details:
No
Agar and Dextrose are used for the preparation of the potato dextrose agar culture medium, needed for maintenance and
culture of Aspergillus niger strains. Prices have been oftained from Aldrich catalogue 2003-2004.
Pyridine and acetic anhydride are needed for the control analysis of citric acid. Prices have been obtained from Aldrich
catalogue 2003-2004.
Macropipettors and tips are needed to speed up analytical work. Prices are from Cole Parmer catalogue 1995-1996.
Aspergillus niger strains are essential to the project. Prices are from Deutsche Sammlung von Mikroorganismen und
Zellkulturen GmbH.