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Glycerol Fermentation of Starch Paul Holland Figard Iowa State College

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Glycerol Fermentation of Starch Paul Holland Figard Iowa State College Iowa State University Capstones, Theses and Retrospective Theses and Dissertations Dissertations 1949 Glycerol fermentation of starch Paul Holland Figard Iowa State College Follow this and additional works at: https://lib.dr.iastate.edu/rtd Part of the Biochemistry Commons Recommended Citation Figard, Paul Holland, "Glycerol fermentation of starch " (1949). Retrospective Theses and Dissertations. 13345. https://lib.dr.iastate.edu/rtd/13345 This Dissertation is brought to you for free and open access by the Iowa State University Capstones, Theses and Dissertations at Iowa State University Digital Repository. It has been accepted for inclusion in Retrospective Theses and Dissertations by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. NOTE TO USERS This reproduction is the best copy available. UMI |-nt> GLYCEROL PERWTATION OP STARCH by Paul H, Pigard A Thesis Submitted to the Gradmte Faculty In Partial Pulfillaent of The RequlreiMinta foi* the Degree of DOCTOR OP PHILOSOPHI Major Subject j Biophysical Ohemist-pj Approved! Signature was redacted for privacy. Signature was redacted for privacy. Head of Major Department Signature was redacted for privacy. Dean of Graduate College Iowa State College 1949 UMI Number: DP12463 INFORMATION TO USERS The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleed-through, substandard margins, and improper alignment can adversely affect reproduction. In the unlikely event that the author did not send a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion. UMI UMI Microform DP12463 Copyright 2005 by ProQuest Information and Learning Company. All rights reserved. This microform edition is protected against unauthorized copying under Title 17, United States Code. ProQuest Information and Learning Company 300 North Zeeb Road P.O. Box 1346 Ann Arbor, Ml 48106-1346 3 a.1 ACKNOWLEDGMENTS fh« author acknowledges his Indebtedness to Dr, Lm k, Ond«rkofler for the suggestion of the problem and for the helpful advice during the course of Its study. V T9/^^ ill TABIE OP COITEMIS AGKIOILEDGMIIIS. ii I,. IimODUCTIOl 1 II. HISfORICAI... ................... 5 III, EXPERIMENTAL. ................. 26 A. Materials. 26 !• Cornstarch. ...... 26 2. Steep water 26- 3. Yeasts, 26 4. Bacterium 27 5. Sulfites. ............... 28 B. Analytical Methods 28 1, Determination of alcohol. ....... 28 2, Determination of sugar. ........ 29' 3, Determination of sulfite. ....... S9 4m Determination of acetaldehyd# ..... 30 5, Determination of glycerol ....... 30 C. An Investigation of Yarlous Aldehyde- fixing Agents To Induce the Glycerol Fermentation of Aeid-hydrolyzed Starch ... 31 D. The Fermentation of Acid-hydrolyzed Cornstarch by Saccharoiaycea cerevisiae in the Presence of Sulfite 33 1. Acid-hydrolysis of cornstarch ..... 33 2. Effect of addition of nutrients to glycerol fermentations ....... 40 3. Studies on the glycerol fermentation of cornstarch with various sulfites . 44 iv Esse 4. Effect of pH on the glycerol yields obtained from acld- hjdrolyzed starch with various sulfites, ............ 55 5, Effect on glycerol yields of the addition of sulfiir dioxide to magnesium sulfite fermentations • 58 6# Effect of varying the mash con­ centration and temperature on the glycerol yields obtained from acld-hydroiyzed starch •mashes. 62 7, Effect on glycerol yield of de­ laying the addition of sulfite to the fermentation 64 8, Effect on glycerol yields of accliiaatlzatlon of yeast to magnesium sulfite 66 9, Effect on glycerol yields of activating the yeast before inoculation 67 1, The Permentation of Acld-hydrolyzed Cornstarch by Bacillus 3ubtllis« » • . 69 1, Effect of mash concentration and nutrients on the yields of glycerol obtained from acid- hydrolyzed starch by Bacillus subtllls fermentations. * . « 69 2, Effect on glycerol yields of adding sulfite to Bacillus aubtilis fermentations. • • • . 70 3«, Effect of pH and stirring on the glycerol yields from Bacillus subtllla fermentations of hydro- lyzed starch, 72 P. The Peraentatlon of Acld-hydrolyzed Cornstarch by ZyKOsaccharomyces acldifaclens . .1 . .• . 73 V Imi 1» Effect of nutrients and sulfite on ZjgoaaGcharomyces fementations. • , ». 73 2. Effect on glycerol yield of pH and the use of msslve inoculations In ZjROB&ccfaaromyces fermentations • , 75 I¥. DISCUSSIOl. 77 V, SUMIttRY AMD CONCLUSIONS 80 ¥1, LITEMTURE CITED, 83 -1- I, iMHiODlJCTIOIf Gljcerolj or glycerine as it is eomraonlj called# has become a r@Tj Importwit Industrial chemioal.. In addi­ tion to the tisss for which it is essential there ar© a wide variety of applications which would lead to expanded eonsuiaption of th© commodity if it were availabl® in larger qtiantltles at a reasonable cost.. At present it is uaad most extensively in textiles^ resins, and ©Kplosives, Th© laat -use was brought to our attention by the household fat salvage project during th© recent war* Tliis conserv­ ing of fats also ©niphaslzed the fact that our principal source of glycerol was the fat-splitting proc#ss» A new synthetic plant which went into operation in 1948 is ex­ pected to assure a more adeqmte supply of pure glycerol. Th© feraentation processes for producing glycerol have been restricted mostly to war-time use when the ne­ cessity for having it for explosive imnufacture overcam© considerations of cost# Germany msde considerable glycerol by fermontation during the World War of 1914-1918., fhelr process mm developed by Conns teln and Iiiidock© and con­ sisted of the ferraentatlon of beet sugar in th© presence of sodium sulfite as a fixing agent for acetftldehyde. By a ratter inefficient method Germany usad the process in -2- 24 faeturlea with the smaller ones shipping the feraentdd slop to th© larger ones for th® recovery of th© glycerol# According to th© description given by Lawrl® (1928) th© output amounted to approximately 1000 tons each month after the factories got into operation on a large seal©# A littl® glycerol was laade by feraentation in ths United Statas aifter th© Genaan proceas had been developed, but h@r« an alkalins method using sodium carbonate was worked out by loff# England nade glycerol by a fermentation process of Cocking and Lilly which waa reported to give very good yields approaching th©;theoretical value* Ihis method was similar to that of Connstein and Liidecke but used sodium bisulflt© in addition to the sodium sulfit# in order to obtain Increased yields of glycerol. In recent years no very drastic changes have been developed in the proceases for producing glycerol by fer­ mentation* Most of the later patents in the field are based on only slight modifications of the proceases men­ tioned above. Much work has been done on the problem of recovering th© glycerol from fermentation residues# Thar© have b©en a number of extraction methods worked out which are claimed to be more efficient than th© older distilla­ tion methods, lacperlenc® has shown that much of th® difficulty and expense involved in the fermentation procaaaea srisea from -3- the TQQoverj of the glycerol. The most troublesome factor is the high concentrations of soluble salts present in tha f©rra@ntod beers. For this reason Hickej (1941) investigated the use of leas soluble sulfites. He employed magnegium or calelum siilflta instead of the sodiua salt». Unfortun­ ately th® yields were not as hlghj however, th© recovei^ of the glycerol should b© aingsler sine© the cftlcima and m&gnmimi salts can b© removed by filtration. Sodium ions cannot b© gotten rid of so ©asily^ for no coistton aodiuia salts are insolublo. Most of th© feraentation procedures for glycerol that are described in th© literature use sugar as the raw ma- terial# but it would b© desirable from th© standpoint of initial cost to use starch* There are very few references to the use of starch for this f©mentation although it is suggested in a few instancea, Connstein and Ludeck© men­ tion the use of a saccharified starch mash in one of their patents, but they give no data on its use,. The theoretical yield of glycerol is approxlma'toly 51 par cent, and thor© are some claims of very nearly this value* However, th® majority of th® yields reported for large-scale fermentations of commercial slse are in th© region between 20 and 30 per cent. If the yields are given for glycerol recovered, they are even lower. There is correlation between the size of th© Inocula and the yields obtained. -4- Ih© purpose of tlie Investigation on which this thesis is based was to determine the best conditions for a glycerol fementation process using starch as the fermentation sub- strat®. It was hoped that a process could bo developed which might ha^e practical application for coiamerclal gljcerol production* ?o this end most of the work was don© with th® use of those sulfites which would not increase the concentration of soluble salts enough to make the glycerol reeovsrj too difficult* 5- II. HISTORICM. ©le history of glycerol from its preparation by Scheele in, 1779 -until 19S8 is very well covered in the monograph ^ Lawi® (1928)« For that reason it will b© necessary to deal here with only those phases of the subject directly concerned with th© present investigation. The principal points of interest will be th© fermentation laethods for the formation of glycerol# Ih© literattare mentions various synthstio methods for preparing glycerol. Wurtz (1857) made it hj reacting l»2#3,-trlbromopropa»e with silver acetate and hydrolyzing the product, triacetln, with alkali* When it was discover­ ed that propylene could b© chlorinated to allyl chloride, it waa realized that this reaction could b« used as an ia- portan.t starting atop for the synthesis of glycerol. The industrial prospects of this method war© discussed by Ii©vey (1938), and he concluded that It was ©eonomically sound# Williams (1941) also evaluated th© economic factors and presented the process as a desirable method for glycerol production.
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