Catalytic transformation of castor cake by Penicillium fungus

Castor cake is a by product from castor seed but remains as a substance with restricted application potential due to its allergenic and toxic properties.People traditionally have treated the castor cake by wild fermentation for the catalytic transformation of anti-metabolites into their corresponding non-toxic forms for improving its utility.

This age-old practice was scientifically approached by the Brazilian researchers where they subjected castor cake to solid state fermentation.Penicillium simplicissimum fungus was chosen as the catalytic factor because of its potency to manipulate in castor cake substrate and generate lipid metabolizing lipase enzymes.Solid state fermentation(dessicated fermentation) was performed for the cultivation of fungus and the products, enzyme and fermented castor cake were partitioned by solid-liquid phase differential extraction.Enzyme lipase magnitude in crude isolated fraction was determined from its activity against synthetic substrate while the debris cake fermented was evaluated for its toxic protein contents and functional activities by immunoassays and immuno sensitive cell culture assays respectively.The results proved to be promising as the toxin titre and allergens induced cell degranulation were seen to be minimum and the microbial enzyme production was noticed to a significant extent as well from specific activity studies.

Results turn out to be promising but the experiment would have been better if an utility for the fermented castor cake in biology was defined.The solid state technology by fungus seems to be a viable one on commodities like castor cake to generate high value products when time factor is not signified to a greater extent.

For reference

Medium formulation in ricin vaccine

Ricin is a noxious cytotoxin present in castor bean.People have long been pursuing research to understand its functional activities,physical structure and quantitative magnitude in beans etc.In spite of such extensive studies, no effective solution has been formulated to ricin intoxication. However researchers have found vaccine therapy as a promising route to combat the toxin.In ricin vaccinology, the wild toxin has been modified in such a way so that it tends to retain its antigenicity to boost the host defence system but not the toxic activity, called it as “ricin toxoid”.Genetic engineering  is a standardized technique to precisely produce variants in ricin and the candidate with expected property are subsequently chosen.

US scientists have identified an elite ricin toxoid from earlier studies and made an evaluation on vaccine formulation to achieve enhanced antigenicity and stability.Initially they treated the protein in phosphate buffer solution and found structural and functional tolerance in protein after cyclic exposures to extreme temperature ranges.To further improve its adjuvant adherence and related antigenicity, toxoid was treated with aluminium hydroxide adjuvant in succinate buffer and evaluated by differential centrifugation and immuno assays.The results indicated that aluminium hydroxide showed a good compatibility with toxoid by co-sedimentation of protein with gel matrix and also elicited a strong immune response, confirmed by antibody titre studies.Functional confirmation did not seem to be significantly affected by conjugation with adjuvant and Fourier transform infrared spectroscopy was applied to understand it.

Scientists in this study have made a complete profile in ricin vaccine formulation and its influence in conferring active immunity to primitive model systems but the confirmation is to be arrived from human clinical trials as extrapolation of data from mice to humans is difficult .

For reference:”ricin+breakth

Mapping of ricin by tools of molecular biology

Ricin toxin from castor seeds has been well established as a cytotoxin interferring with ribosomal functions.The activity of any protein generally is conferred by the domains present in it. This made scientists to probe the domains and  critical amino acids for the cytotoxic activity of ricin. Researchers in USA have made molecular analysis of castor bean toxin at the genetic level where tools of genetic engineering are exploited to address their domain mapping studies.The experimentation involved synthetic mutation process in genetic determinants of ricin protein for hypothetical amino acid, glutamate to alanine which basically is distant to the former constituent in terms of its physio-chemical properties and makes it an ideal substituent to understand the magnitude of glutamate in the ricin function.Their data in the end from mutant protein’s activity indicated that the target glutamate is not of primary significance in ricin’ toxicological function.

Domain mapping by mutations is an iterative process but an intense technique to make a complete scrutiny of the sub components in a complex molecule and their implications to the ultimate emergence of functions from bulk.

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Interpretation of castor bean and Ricin as reproductive toxins.

Ricin toxin is well to act as a sub- acute toxin in humans to produce lethality between 2-5 days on the basis of concentration and portals of administration of toxin.However its role in reproductive toxicity has not been documented well and Jordanian scientists have made an attempt to assess the role for ricin in reproductive toxicity in pregnant female rats.

Rats intraperitoneally treated with ricin and castor bean extracts displayed symptoms of body weight loss and death of foetuses at 5-9 days of pregnancy while attrition of implantation sites was noticed with first 6 consecutive days of pregnancy.The toxicological function was not significantly reversed with gonadotrophin hormone in experimental subjects exposed to toxin.Hence the researchers were able to conclude that castor bean constituents including ricin have anti implantation and ovulation properties through their overlapped reproductive toxicological studies.

Results of this study is difficult to interpret as multiple parameters including strain,species,sex,age,feed,time,concentration of the toxin,mode of administration,animal size of the experimental study have a profound influence in the outcome of the study.Adopting International standard guidelines in toxicology  will greatly improves the precision and accuracy of the experiment.

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Role for MALDI-MS in ricin detection.

Ricin is a lethal cytotoxin present abundantly in castor seed.People have identified the protein way back but no effective, timely sensitive antidote or assay kit has ever been formulated so far.However advancements in protein technology have rendered a solution today through application of an advanced mass spectrometry for the detection of ricin, called as Matrix Assisted Laser Desorption Ionization Mass spectrometry(MALDI-MS).It is extensively applied for detection of proteins which are unknown and whose magnitude is difficult to determine by conventional resolution principles.

Australian scientists tried to exploit it in the context of ricin chemistry and have prepared a polymorphic peptide mass fingerprint of ricin and its related proteins  following an initial treatment with trypsin.They were excited to watch the extent of protein resolution by MALDI-MS in purified and crude preparations of ricin from castor seeds without ambiguity. Scientists further claimed that their protocol has  scope for improvements and consumes minimal time to probe the presence of toxin from sample preparations.

MALDI-MS proved its utility in toxin chemistry of proteins as a valuable tool and will expand its wing to conquer new protein niches in biology.

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Valuable metabolite probe from castor meal

Castor meal is a concentrated residue matter produced after extraction of oil from castor seeds.Its application is so restricted due to ricin and poor molecular biochemical knowledge about the material.A surprising breakthrough has been made by the discovery of galactinol, a carbohydrate derivative associated with raffinose metabolism in plants to an appreciable extent on mass basis.

Experimental designs including chromatography and mass spectrometry were applied to isolate and confirm the purity of galactinol in castor meal.The scientists further stated that it is exciting as research in past in the concerned field is hindered by a lack of availability of commercial source of galactinol.

Galactinol discovery in castor meal illuminates the potential of castor meal which is currently applied as a field manure in most of the castor growing regions.Identification of high value metabolites would tremendously add the commercial value to this product on an international scale.

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Processed castor bean meal as a probable feed for poultry.

Castor bean meal(CBM) is well known for its toxicity functions due to the presence of ricin protein and other castor plant based metabolites.A classical but a productive research study from the Rutgers University,USA revealed a complete, strange fact about castor bean cake.Researchers Laufey Vilhjalmsdottir and Hans Fischer have chosen CBM and treated it  with warm water to exclude its toxic debris materials.They later fed the processed material to the bred chicks as a test proteinaceous diet to assess an increase in biomass and were able to notice not alone a loss of toxicity from the fed material but also an incrementation in the mass development of experiemental subjects which enabled scientists to understand the scope of CBM as a potent future, feedstock in poultry.The results also revealed that CBM has to be supplemented along with some essential amino acids to achieve a superior activity in chicks which otherwise proved to be inferior performing to soybean diet.

In this analytical study, scientists were able to prove the efficacy of the deoiled cake from castor but the experimental robustness with appropriate controls and parameters are necessary to confirm the compatibility of CBM as a routine feed for the poultry and other livestocks.

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Increase in Freight Rate Will Make the Castor De-oiled Cake Production Unviable?

In India, the ministry under Mamata Banerjee has re-classified de-oiled cakes and oil meals from Class 110 to Class 120, effective December 27,2010 which will entail a freight rate increase between 11 and 13 per cent. Since oil cake and meals are used effectively as high nutrient bird and animal feeds, the rise in raw material prices will be passed on to consumers.

Solvent Extractors Association (SEA), in a memorandum to the railways minister and chairman of the Railway Board as well as secretary, Ministry of Agriculture, Commerce, Animal Husbandry and Finance, highlighted that de-oiled cakes were locally produced, traded and occasionally exported on wafer-thin margins. This increase in freight rate would make the production of de-oiled cakes unviable thereby forcing many processing plants to decrease their production and thereafter leading to
De-oiled cakes end-use is in manufacture of compound feeds for cattle, poultry and aqua and also as natural fertilisers and hence such a large increase in freight would place great constraint on the viability of dairy and food industry as well as on the farmers. Also, exports of de-oiled cakes, which face stiff international competition from giants in the US and South America, will be adversely affected. In fact, to compete in the international market, the government has granted a 2 per cent incentive under the Focus Product Scheme for exports of some de-oiled cakes and 5 per cent under Vishesh Krishi Gram Upaj Yojana for castor de-oiled cake. The hike in freight will totally nullify the benefit granted to the exporters of de-oiled cakes and would place them at a great disadvantage.