Verfügbarkeit: Purification and characterization of LOX isoenzymes from germinating barley

Purification and characterization of LOX isoenzymes from germinating barley: biotransformation of complex lipids

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Bibliographische Detailangaben
Beteilige Person:	Almeida, Roberto Barbosa de (VerfasserIn)
Format:	Buch
Sprache:	Englisch
Veröffentlicht:	2005
Schlagwörter:	Hochschulschrift
Links:	http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=014639598&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA
Beschreibung:	Berlin, Techn. Univ., Diss., 2005
Umfang:	XIII, 121 S. Ill., graph. Darst.

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Datensatz im Suchindex

DE-BY-TUM_call_number	0001 DA 2006 28
DE-BY-TUM_katkey	1546615
DE-BY-TUM_location	Mag
DE-BY-TUM_media_number	040006150874
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adam_text	PURIFICATION AND CHARACTERIZATION OF LOX ISOENZYMES FROM GERMINATING BARLEY : BIOTRANSFORMATION OF COMPLEX LIPIDS VORGELEGT VON M.SC. ROBERTO BARBOSA DE ALMEIDA AUS LORENA, SAO PAULO, BRASILIEN VON DER FAKULTAT III - PROZESSWISSENSCHAFTEN DER TECHNISCHEN UNIVERSITAT BERLIN ZUR ERIANGUNG DES AKADEMISCHEN GRADES DOKTOR DER INGENIEURWISSENSCHAFTEN - DR.-LNG. - GENEHMIGTE DISSERTATION PROMOTIONSAUSSCHUSS: VORSITZENDER: PROF. DR.-LNG. DR. E.H. FRIEDRICH MEUSER BERICHTER: PROF. DR. RER. NAT. ROLAND TRESSL BERICHTER: PROF. EM. DR.-LNG. KARL WACKERBAUER TAG DER WISSENSCHAFTIICHEN AUSSPRACHE : 11. AUGUST 2005 BERLIN 2005 D83 INDEX 1 - INTRODUCTION 1 1.1 - FLAVOUR STABILITY 2 1.2 - LIPIDS IN THE BREWING PROCESS 6 1.3 - OXIDATION OF FATTY ACIDS 7 1.3.1 - PHOTOOXIDATION OF FATTY ACIDS 7 1.3.2 - AUTOXIDATION OF FATTY ACIDS 9 1.3.3 - ENZYMATIC OXIDATION OF FATTY ACIDS 10 1.3.3.1 - LIPOXYGENASES 11 1.3.3.2 - LOX IN THE MALTING / BEER PROCESS 12 1.3.3.3 - LIPOXYGENASE - MECHANISM 14 1.3.3.4 - LOX - SUBSTRATE ACCEPTANCE 16 1.3.3.5 - HYDROXY FATTY ACIDS IN THE MALTING / BEER PROCESS 18 1.3.3.6 - HYDROPEROXY FATTY ACID METABOLIZING ENZYMES (HPME) 20 2 - AIM S OF THE THESIS 23 3 - MATERIALS AND METHODS 24 3.1 - CHEMICALS 24 3.2 - RAW MATERIAL 24 3.3 - SYNTHESIS 25 3.3.1 - SYNTHESIS OF (S)-9-HYDROPEROXY-(E,Z)-10,12-OCTADECADIENOIC ACID 25 3.3.1.1 - FIRST STEP - EXTRACTION OF LIPOXYGENASE FROM TOMATO 25 3.3.1.2 - SECOND STEP - ENZYMATIC TRANSFORMATION OF LINOLEIC ACID 25 3.3.2 - SYNTHESIS OF (S)-9-HYDROXY-(E,Z)-10,12-OCTADECADIENOIC ACID 26 3.3.3 - SYNTHESIS OF 9-KETO-(E,Z)-10,12-OCTADECADIENOIC ACID 26 3.3.4 - SYNTHESIS OF RACEMIC 9-HYDROXY-(E,Z)-10,12-OCTADECADIENOIC ACID 27 3.3.5 - SYNTHESIS OF (S)-13-HYDROPEROXY-(Z,E)-9,11-OCTADECADIENOIC ACID 28 3.3.6 - SYNTHESIS OF (S)-13-HYDROXY-(Z,E)-9,11-OCTADECADIENOIC ACID 28 3.3.7 - SYNTHESIS OF 13-KETO-(Z,E)-9,11-OCTADECADIENOIC ACID 28 3.3.8 - SYNTHESIS OF RACEMIC 13-HYDROXY-(Z,E)-9,11-OCTADECADIENOIC ACID 28 3.4 - ENZYME ASSAYS 29 3.4.1 - LIPOXYGENASE ASSAY 29 3.4.2 - HYDROPEROXY - METABOLIZING ENZYMES (HPME) ASSAY 30 3.5 - LIPOXYGENASE ISOENZYMES - EXTRACTION AND PARTIAL PURIFICATION / SEPARATION 30 3.5.1 - LIPOXYGENASES - EXTRACTION AND PARTIAL PURIFICATION 30 3.5.2 - LIPOXYGENASES - ISOENZYMES PARTIAL PURIFICATION / SEPARATION 31 3.5.2.1 - HYDROXYLAPATITE CHROMATOGRAPHY 31 3.5.2.2 - ISOELECTRIC FOCUSSING (IEF) 31 3.6 - LOX ISOENZYMES - CHARACTERIZATION 32 3.6.1 - SDS-PAGE 33 3.6.2 - PH OPTIMUM 33 3.6.3 - KINETIC PARAMETERS 33 3.6.4 - INFLUENCE OF TEMPERATURE ON THE ENZYME ACTIVITY 34 3.6.5 - PROTEIN CONCENTRATION 34 3.7 - EXTRACTION OF MALT LIPIDS - FATTY ACID COMPOSITION 34 3.7.1 - EXTRACTION OF NON-POLAR LIPIDS 34 3.7.2 - EXTRACTION OF POLAR LIPIDS 35 3.7.3 - DETERMINATION OF THE FATTY ACID COMPOSITION OF THE EXTRACTED MALT LIPIDS 35 3.8 - PREPARATION OF STOCK EMULSIONS 36 3.8.1- SUBSTRATE STOCK SOLUTION OF FREE LINOLEIC ACID 36 3.8.2 - SUBSTRATE STOCK EMULSIONS OF ESTERIFIED DERIVATIVES OF LINOLEIC ACID 36 3.8.3 - SUBSTRATE STOCK EMULSIONS OF EXTRACTED LIPIDS 36 3.9 - PREPARATIVE ENZYMATIC TRANSFORMATIONS 37 3.9.1 - ENZYMATIC TRANSFORMATIONS USING FREE LINOLEIC ACID AS SUBSTRATE 37 3.9.1.1 - HYDROXYLAPATITE CHROMATOGRAPHY PARTIAL PURIFIED LOX 37 3.9.1.2 - ISOELECTRIC FOCUSSING PARTIAL PURIFIED LOX 37 3.9.1.3 - CRUDE EXTRACT LOX 37 IV 3.9.2 - ENZYMATIC TRANSFORMATIONS USING ESTERIFIED DERIVATIVES OF LINOLEIC ACID AS SUBSTRATE 38 3.9.2.1 - ISOELECTRIC FOCUSSING PARTIAL PURIFIED LOX 38 3.9.2.2 - CRUDE EXTRACT LOX 38 3.9.3 - ENZYMATIC TRANSFORMATIONS USING EXTRACTED LIPIDS AS SUBSTRATE 38 3.9.3.1 - ISOELECTRIC FOCUSSING PARTIAL PURIFIED LOX 38 3.10 - PRODUCT ANALYSIS OF THE ENZYMATIC TRANSFORMATIONS 38 3.10.1 - DETERMINATION OF THE AMOUNT OF THE REGIOISOMERS 9- AND 13-HPODE AND THE REMAINED LINOLEIC ACID. 38 3.10.1.1 -SAMPLE PREPARATION 38 3.10.1.2 -GC- MS ANALYSIS 40 3.10.2 - DETERMINATION OF THE STEREOISOMER RATIOS FROM 9-HPODE AND 13-HPODE 41 3.10.2.1-SAMPLE PREPARATION 41 3.10.2.2-HPLC-ANALYSIS 41 3.11 - DETERMINATION OF THE LOXS RELATIVE ACTIVITIES 42 3.12 - STATISTIC METHODOLOGY 43 4 - PURIFICATION, SEPARATION AND CHARACTERIZATION OF LOX ISOENZYMES USING FREE LINOLEIC ACID * 44 4.1 - INTRODUCTION 44 4.2 - RESULTS 45 4.2.1 - ANALYSIS OF 9- AND 13-HPODE 45 4.2.2 - PURIFICATION / SEPARATION METHODS 50 4.2.2.1 - HYDROXYLAPATITE CHROMATOGRAPHY 50 4.2.2.2 - ISOELECTRIC FOCUSSING 52 4.2.3 - REGIQSELECTIVITY OF LOX ISOENZYMES WITH FREE LINOLEIC ACID AS SUBSTRATE. 53 4.2.3.1 - HYDROXYLAPATITE CHROMATOGRAPHY 53 4.2.3.2 - ISOELECTRIC FOCUSSING 54 4.2.4 - TOTAL AMOUNT OF HPODE AFTER ENZYMATIC TRANSFORMATIONS OF LINOLEIC ACID WITH IEF SEPARATED LOX 55 4.2.5 * HYDROPEROXIDE-METABOLISING ENZYMES 58 4.2.6 - STEREOSELECTIVITY OF PARTIALLY PURIFIED LOX ISOENZYMES 61 4.2.7 - FORMATION OF (E.E)-ISOMERS 62 4.2.8 - KINETIC PARAMETERS 63 4.2.9 - ELECTROPHORESIS 66 4.2.10-PH OPTIMUM 66 4.2.11 - INFLUENCE OF TEMPERATURE 68 4.3 - DISCUSSION 69 4.4 - SUMMARY 72 5 - SUBSTRATE-, REGIO- AND STEREOSPECIFICITY OF LOX ISOENZYMES WITH COMPLEX LIPIDS AND EXTRACTED MALT LIPIDS 74 5.1 -INTRODUCTION 74 5.2 - RESULTS 75 5.2.1 - ANALYSIS OF EXTRACTED LIPIDS 75 5.2.2 - TRANSFORMATION OF TRILINOLEIN WITH CRUDE ENZYME EXTRACT 77 5.2.3 - ENZYMATIC TRANSFORMATION OF ESTERIFIED FATTY ACIDS BY LOX1 AND LOX2 77 5.2.4 - RELATIVE ACTIVITIES OF LOX ISOENZYMES WITH FREE AND ESTERIFIED LINOLEIC ACIDS 79 5.2.5 - REGIOSELECTIVITY OF LOX ISOENZYMES WITH ESTERIFIED LINOLEIC ACID 80 5.2.6 - STEREOSELECTIVITY OF LOX ISOENZYMES WITH FREE AND ESTERIFIED DERIVATIVES OF LINOLEIC ACID 83 5.2.7 - FATTY ACID COMPOSITION OF POLAR LIPIDS AND NON-POLAR LIPIDS EXTRACTED FROM MALT 85 5.2.8 - REGIOSELECTIVITY OF LOX ISOENZYMES WITH MALT LIPID EXTRACTS 86 5.2.9 - STEREOSELECTIVITY OF LOX ISOENZYMES WITH MALT LIPID EXTRACTS 87 5.3 - DISCUSSION 89 5.3.1 - BIOTRANSFORMATION OF ESTERIFIED LINOLEIC ACIDS WITH LOX1 89 5.3.2 - BIOTRANSFORMATION OF ESTERIFIED LINOLEIC ACIDS WITH LOX2 90 5.3.3 - BIOTRANSFORMATION OF TRILINOLEIN WITH CRUDE ENZYME EXTRACT 91 5.3.4 - BIOTRANSFORMATION OF EXTRACTED POLAR LIPIDS WITH LOX1 AND LOX2 92 5.3.5 - BIOTRANSFORMATION OF EXTRACTED NON-POLAR LIPIDS WITH LOX1 AND LOX2 93 5.4 - SUMMARY 95 VI 6 - CONCLUSION 97 7- REFERENCES 99 8-APPENDIX 116 VII
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author	Almeida, Roberto Barbosa de
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dewey-ones	572 - Biochemistry
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spellingShingle	Almeida, Roberto Barbosa de Purification and characterization of LOX isoenzymes from germinating barley biotransformation of complex lipids
subject_GND	(DE-588)4113937-9
title	Purification and characterization of LOX isoenzymes from germinating barley biotransformation of complex lipids
title_auth	Purification and characterization of LOX isoenzymes from germinating barley biotransformation of complex lipids
title_exact_search	Purification and characterization of LOX isoenzymes from germinating barley biotransformation of complex lipids
title_full	Purification and characterization of LOX isoenzymes from germinating barley biotransformation of complex lipids von Roberto Barbosa de Almeida
title_fullStr	Purification and characterization of LOX isoenzymes from germinating barley biotransformation of complex lipids von Roberto Barbosa de Almeida
title_full_unstemmed	Purification and characterization of LOX isoenzymes from germinating barley biotransformation of complex lipids von Roberto Barbosa de Almeida
title_short	Purification and characterization of LOX isoenzymes from germinating barley
title_sort	purification and characterization of lox isoenzymes from germinating barley biotransformation of complex lipids
title_sub	biotransformation of complex lipids
topic_facet	Hochschulschrift
url	http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=014639598&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA
work_keys_str_mv	AT almeidarobertobarbosade purificationandcharacterizationofloxisoenzymesfromgerminatingbarleybiotransformationofcomplexlipids

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