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    29 views31 pages

    Pectin Extraction Form Coffee Pulp

    Uploaded by

    Thi Ngoc Dinh Ho

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    © All Rights Reserved
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    «2 United States Patent Belaleazar Otalora US 9,896,572 B2 Feb. 20, 2018 ‘US009896572B2 (10) Patent No.: (4s) Date of Patent: (S4) PECTIN EXTRACTION FROM COFFEE. PULP, oy @ Applicant: Pectoof BLV,, AB Wageaingen (NL) Inventor: Andres Felipe Belaleazar Otalora, CE Amsterdam (NL) 03) © Assignee: Peeteof B.V;, Wageningen (NL) Notice: Subject to any disclaimer, the term ofthis patent is extended or adjusted under 35 USC. 1540) by 0 days an @y (86) Appl.No: 14(647,569 PCT Filed: Nov. 27, 2013 PCT No. 5.371 XD, (2) Date: PCTEPZO1S078811 May 27, 2015 (87) PCT Pub. Nos WO2014/083032 PCT Pub, Date: Fun, 5,20 Prior Publication Data US 2015/0307634 Al Oct. 29, 2015 ws) G0) Foreign Application Priority Data Nov. 28, 2012 (EP) 12194850 (1) meer. cos S06 cose 3700 C12? 19708 ADF S16 Ch2P 1948 Us. cL cbc (2006.01) (2006.01), (2006.01) (2006.01), (2006.01) 2) COBL 5/06 (2013.01); A23F 5/168, (2013.01): 423F §/166 2013.01); COB 37/0045 (2013.01): COBB 37/0048 (2013.01); CRP 19/0 2013.01}; C12P 19444 (2013.01): CRY IMT 201301); C13¥ 30100 (201301) (58) Fleld of Classitication Search CPC COSI. 5/05; CORB 37/0048; COS 37/0088: C12P 19108; C12P 19/48; CI2Y LILOL; (C12Y 301/00; A23P 5/163; AMP S/166 See application file for complete search history: 66) References Cited USS. PATENT DOCUMENTS. 61987 Rombout 2001. Badofeen soa. a (ol Bie cost 370045 2488 20080220485 AL 11/2003 Ni FOREIGN PATENT DOCUMENTS wossonas0 Woon zTH8 wouitorss22 wo wo wo 21996 52000 eon (OTHER PUBLICATIONS (Garcia etal. “Characterization of Cotes Posi ‘Wiss u-Technl, vol. 24: 125-129 Potetial alternative uses of cole wastes an hy-proet(2005) Interntional Coffe Organization. ED 196708: 1-4 ‘Buetiolt et al, “Preparation and Properties of Enzjmatially and Chemically Mitied Sugar Beet Pectins.”Seiene Dies, Cabo Ihre Polymers 88,2004, pp. 149-161 Calle, “Methods De Exuatcion De Las Pectinas Del. Café" CCencaf, Colombia, vol 12, No.2 1962 pp. 69-74 Esquivel a, "Functional Propeties of Coffe and Coffs ty- prc Food Research International, 46,2012, pp. 488-495, Fissore eal, “Rheological Performance of Petin-eniched Pro ‘ucts Iolated from Red Boot (Beta vulgris L. var condita) through Alkaline and Enzymatic Treatments" Food Hiydiocolids, 26, 2012. pp. 249-260, Usodsiks cal, “Pooveatments to Eahance the Digestibiliy of ignoellulosc Biomass,” Bioresource Techtolog. 100, 2009p, ots, Instituto Centoamericano De Investigacion ¥ Tecnologia Inds tra, “Indostrializtion of Colas Pectin” un. 1986, 56 pages setieved from URL: pl ssid gov'pd’ does PDAAWSI7 pall Jun. 24,201 ‘MeMamis et al, “Polyphenol Interactions. Part 1. Inroductio ‘Some Observations om the Reveruible Complexation of Polypienl ‘vith Proteins and Polysachardes.” J, Chem Soc, Perkin Tras. 1985, pp. L918, Muri, eal, “EMeet of Bisulfite Addition om the Chemical Composition and Cellule Content Fractions of Dehyrted Coles Pulp! J. gic. Food Chem, 28, No, 1977, pp. 1090-1002, Oowtcrveld et al, “Fovnution of Feri Ack! Dehydiodiers through Oxidative Crosliking of Sugar Beet Pectin” Carboy rate Research 300, 1997, pp. 179-181 ‘Rombouts cal, "Feruloysted Pete Substances fiom Sugar-Beet Pulp Carbohydrate Resear, 154, 1986, pp. 177-187 Sehmelier cal. "Enzymasic Mosiction of Postne and the Int on thes Rheological Properties” Carbohydrate Polymers, 472002, pp. 99-108, * (1991) Lebensm (Continved) Primary Examiner — Robert Yamasski Assistant Examiner — Teresa B Knight (74) Attorney, Agent, or Firm — Ocehiuti & Robliek LLP on ABSTRACT The invention provides golTee pulp treatment process comprising (a) Providing coffee pulp, obtainable from production process for producing green collee beans Irom toffee cherries; (b) extrcting from the coffee pulp pectin ‘comprising extract, wherein extruetion is performed under acid conditions or alkaline conditions, to provide the pectin comprising extract; (¢) enzymatic treatment of the pectin ‘comprising extract, wherein the enzymatic treatment com- prises treatment with one or more enzymes selected fr the group consisting of an esterase and a reductase, 10 provide & enzymatically treated pectin material; and (@) extraction of polyphenol functionalized coffee pectin extract ‘rom the enzymatically eated peetin material 16 Claims, 9 Drawing Sheets US 9,896,572 B2 Page 2 66) References Cited (OTHER PUBLICATIONS ‘Statford, tal, “Inhibition of Spilage Mou Conia by Acetic Acid and Sorbie Acid Involves Dillerent Modes of Aston, Rei ing Modification ofthe Clasical Weak Theory” International Journal of Food Microbiology. 136, 2009, pp. 37-8. Wehr ta, "Alkali Hydroxie-indaced Galation of Pein” Food Hyrlocolloids, 18,2008, pp. 375-378 * cited by examiner U.S. Patent Feb. 20, 2018 Sheet 1 of 9 US 9,896,572 B2 coe r 100 200 300 10/110 é 210 320 | +f | 2 310 | ¥ 400-| 410—~ | y a 500-7 | me -~600 5107} ad [| 610 ¥ 700 J - aaa 710 \ 810 800 FIG. 1 U.S. Patent Feb. 20, 2018 Sheet 2 of 9 US 9,896,572 B2 U.S. Patent Feb. 20, 2018 Sheet 3 of 9 US 9,896,572 B2 FIG. 2b U.S. Patent Feb. 20, 2018 Sheet 4 of 9 US 9,896,572 B2 U.S. Patent Feb. 20, 2018 Sheet 5 of 9 US 9,896,572 B2 U.S. Patent Feb. 20, 2018 Sheet 6 of 9 US 9,896,572 B2 FIG. 5a U.S. Patent Feb. 20, 2018 Sheet 7 of 9 US 9,896,572 B2 U.S. Patent Feb. 20, 2018 Sheet 8 of 9 US 9,896,572 B2 0.60 0.50 0.40 0.30 0.20 0.10 0.00 0 AU 212 2.19 2 — 2.20 + lo 14 — 2.214 FIG. 5d U.S. Patent Feb. 20, 2018 Sheet 9 of 9 US 9,896,572 B2 31 3.2 ~~ 5.20 — 5.21 3.5 Ud 33 L_o; 194 oy |} _j J L. 3.23 3.64 : 5.24 i 35.25 3.12 53.14 38 3.27 3.45 | 3.32 3 161 3.10 588 5.36 rm ) 5.37 z ao |+ 3.39 F340 42 JH [3.41 US 9,896,572 B2 1 PECTIN EXTRACTION FROM COFFEE. PULP, CROSS REFERENCE TO RELATED "APPLICATION ‘This application i the national phase under 35 USC 371 ‘of international application no. PCT/EP2013/074811, fled Nox. 27, 2013, whieh claims the benefit of the procty date ‘of European application no, 12194550.5, filed Nov. 28, 2012. The contents of the aforementioned applications are Jncomporated herein in their entirety. FIELD OF THE INVENTION The invention relates to a coffe pulp treatment process as well as o the product(s) obtained with such process. BACKGROUND OF THE INVENTION With the production of coffe, cof pulp is produced Regardless ofthe source (wet or dey processing) by-prod- ucts and waste products ae olten problematic. For example, pulp and mucilage are relatively acidic, comosive to equip: ‘ment, and dificult to safely dispose. Furthermore, where the pulp is discarded in a landfill or other disposal site, rotting pulp may lead to unpleasant smells, Therefore, by-products ‘and waste products have ofien been viewed as materials, ‘which are either unusable, hazardous, or of negligible valve 'W02004098320 describes a method for isolating a ntsi= ‘ent from coffe cherries or lor producing a food produc that comprises a colfee cherry or portion thereof. In W02004008320, iis particularly preferred that coffee cher- ries will have an extremely low concentration of myeotox- ins, including variows afftoxins, fumonisins, ochratoxins, and/or vomitoxin (DON, deoxynivaleno)), SUMMARY OF THE INVENTION As will be clear from the above, and as also further indicated below; there is a desire to make the coffee pro- ‘duetion process greener, especially by an economic reuse of parts ofthe by-products of the proces, such as colle pulp. In the process to obtain green coffee beans, a major stream ‘of biomass is produced. This biomass is rich in usefil bio ‘compounds; however, a technology really commercially ready for the recovery and use of these compounds is not available, Moreover, the high amounts of toxie compounds (mostly polyphenols and eaffeine) in the streams make the biomass an environmental problem in the coffee producing regions, First some general comments are given below. “After collection ofthe eae cherries the cote cherries ‘are subjected to various processes to oblain the green collec bean (i.e. non-roastad cole bean). Most ofthe world eoflce production is processed in two ways: the dry method and the ‘et mith. In the Wet method, the cheries are collected and pulped fresh, leaving the mucilage (endocarp) and the silver skin attach to the beans; affer pulping the beans go to fermentae tion tanks for period in general in the range of 12-24 hin which the mucilage is released from the beans and silver skin. The beans then are dried (sun or hot air dried), the silver skin is removed and the green beans are packed and stored for further trade. During these processes about 45% ‘ofthe coffee cherry biomass is discarded as waste material, “This biomass has high content of polyphenols and ealleine, ‘and therefore becomes toxic in high concentrations 2 Although composting is an altematve, big extensions of land and hard labor are necessary. On top ofthese requre- ‘meats, the high concentration of polyphenols makes of the tse of this compost poor fertilizer with the risk of poisoning the soil and making it acide, ‘ence, the term “coffee pulp” herein especially refers to the pulp obtained after chery processing. Therefore, the term “coffee pulp” might also reler to “coffee chery pulp Furr, the term “coffee pulp” may also include discarded unripe and overripe cherries, not usable in the production of (high) quality green beans. lence, the tenn coffe pulp may especially refer to one or more of pulp obtained after cheery processing, discarded unripe cheries not usable in the production of (high) quality green beans and discarded ‘verripe cherries not usable in the produetion of (high) quality green beans. The coffee pulp may relate to one or ‘more of the exocarp outer mesocarp (the pp ise), and the :mesocarp (mucilage or parchment) The hull (also known as silver skin or parchment) isnot taken into aecount Note that thesilver skin is part of the seed, not part of the pericarp. The pericarp is especially composed ofthe skin (exoearp). lp (outer mesocarp),mucilage (endocarp). The silver skin is part of the endospemn, It is further referred to amongst thers Esquivel etal, “Functional properties of coffee and coffee by-products”, Food Research Intemational 6 (2012) 488-495, which is incorporated herein by reference. Espe- cially, the coffee pulp is a by-product from the wet method processing, ora semi-dry processing, of eof beans Tn the dry method, the eof cheries ae dried, especially under the sun, for—in general—several days, Therealte, the trial pulp is separated from the green beans by pulping. This tha does not easily allow control of the drying process And may therefore generate a low(er) quality coffee ‘Most ofthe coffee nowadays is processed using the wet method (see above) with variations to lower water consump- ‘ion and contro! over the drying process (semi-dry, natural pulping etc). Nevertheless, relative large amounts of bie- mass are still dumped into rivers, "Nowadays, most ofthe cof pal goes without treatment iretly to huge waste disposal sites without any treatment, eventually toxie compounds from the fermenting cheaics Teach to the rivers, polluting the sources of water in the calles producing regions, Coffee pulp therefore poses serious environmental problem, and is a challenge to the Sustainability of the collee supply chain. With the actual production of coffee reaching 10 million tons per year, Technology to exploit this waste stream is necessary. Cur- rently cole pulp isin genera used only as compost. There thas been reseirch in the use of coffee pip a8 feed for dairy catle in Central and South Americas. Nevertheless, these practices use only a small percentage of the whole Stream due to the anti-nutitional and toxic compounds in the biomass. Purr it has been suggested inthe art to use cede fibers from coffee pulp as well as other sub products from this biomass. However, there is no know teelinology in the extraction separation and modification of pectin from coffee pulp and mucilage Coffee pulp represents 45% of the total weight of the coffee cherry: The pulp biomass is rich in carbohydrates, polyphenols and caffeine. Because the high contents of ‘onganie acids, cathechins, and tannins, the collee pulp and process water pose a serious environmental problem in the regions where production takes place, Coffee discarded streams (the pulp and process water used to soparate the ‘mucilage from the bean in the wet milling factories) have a high BOD (biochemical oxygen demand), which threatens ater sources. One of the components of coffee pulp is US 9,896,572 B2 3 pectin, Pectin is amongst others know inthe Food industry as gelling agent. However, pectin fom collee pulp has bees reported a 2 poor yelling agent and therefore not useful in ood and pharmaceutical applications, Ii theorized thatthe poor gelling properties area result ofthe short length of the postin backbone, the low molecular weight ofthe pectin and the high degree of setylaton ofthe native pectin inthe pp and mucilage of coffee cherry. ‘Hence, it is an aspect of the invention to provide an alterative coflze pulp treatment process, which preferably Jurther at least parly obviate one or more of above-de- scribed drawbacks, Its furder an aspect ofthe invention to provide an alternative pectin, derived from coffee pulp, that ‘canbe used in food applications as gelling agent andr that may have other useful applications. Its furlher an aspect of the invention to provide a solution tothe coffee pulp, by ‘which the coffee production process can. become environ- mentally more sustainable ‘The preseat invention includes the extraction and use of ‘atest one compound extracted from the pulp and muciloge aller depulping and washing of the bean n the wet oF semi ‘dry process of green coffee production. Advantageously, the ‘exinction ofthis bio compotund will reduce concentration of ‘oxic compounds in the processing water of eoffee de- pulping facilities. Further, the extracted compounds in Which pectin is the main component, is high value ingse- «dient forthe food and/or pharmaceutical industries. Further, it surprisingly appears that the extracted bio compounds show the possibilty to be tailored for specifi purposes dae to the diversity of polysaccharides contained inthe extracted postin fraction. The pectin obtained with the process of the invention may allow applications lke as prebotic as well as gelling agents, but also as mesh for sufgcal implants are among the possible ses ofthe compounds extracted accont- ing to the invention, Further the pectin oblained may be used as thickener or emulsifier "The technology suggested here sims forthe extraction of pectin from coffee pulp, and optionally modifiation (i. Junetionalization) of the (extracted) pectin with enzymes Such: modification may include demethylation andor eross- linking the pects thrgh the esterified groups. The toch- nology’ presented here especially sims forthe extraction of pectin from collee pulp, and modification ofthe same pectin with enzymes, to crosslink the pectns through the esterified proups. The enzymatic modifications surprisingly appear to Improve the hyo colloidal properties of the extracted pectin, The properties of the resulting pectin are very attractive, In the process, the remaining waste steam may. ‘amongst others be detoxified through the bydrolization of tannins, polymerization of phenols and removal of caffeine ‘during the process; this will leave the streams with 2 substantially reduced BOD (biological oxygen demand) and ‘COD (chemical oxygen demand). Therefore, the environ- ‘mental impact of coffee production will diminish. ‘The ‘approach may also generate income from the exploitation of the biomass waste as a by-product ofthe coffee chain. The ‘current invention may significantly contribute to improve the sustainability of a major global commodity. Hence, the ‘invention provides a bioretinery approach in which piven ‘chemistry and biotechnology is applied. The process steps ‘may’ coasist of preservation of coffee pulp atthe country of production, shipment to a processing site, separation and Purification ofthe products, and commereatization of these products in their perspective markets. A market may be the market of food ingredients, wherein high quality pectin as a potential replacement of Arabic Gum is sigzested. With the Present innovative technology the coffee pectin can be 0 o 4 tailored to meet the standards of diferent types of appica- tions in the food and pharmaceutical industry. The caffeine content in the remaining waste may advantageously be below 10 ppm, such as even below 1 ppm. Hence, the removal of ealfeine may be very efficient while on the other hand also useful pectin prodict is provided Its known that polyphenols in high concentration can be toxic to cattle inhibit fermentation and growth of microor saanism. Advantageously, in the disclosed invention the presence of polyphenols is actually desited to allow the ‘medication of the pectin without destroying the biopoly- ser. The teclinology i the best alleraive atthe momert, {forthe management and exploitation of coffee waste, There Tore, the disclosed technology might be adopted at a big scale Hence in firs aspect, the invention provides a coffee pulp treatment process comprising: 8, Providing coffee pulp, obtainable from s production process for producing green (jc. non-roasted) collee beans from coffee cherries: b. Eximeting from the colles pulp & pectin comprising ‘extrac, wherein extraction is performed under acid con- ditions or alkaline conditions (or one after the other), 10 provide (or produce) the pectin comprising extract, espe cially wherein the extraction comprises extrcting from the coffee pulp a pectin comprising extract, wherein ‘extraction is performed under (at least acid conditions (optionally) enzymatic twatmeat ofthe pectin comprising extract, wherein the (optional) enzymatic treatment com prises ® treatment with one or more enzymes selected from the group consisting ofan esterase andor a redue> tase, to provide an enzymatically treated (or modified) pectin material, especially polyphenol functionalized eof ee pectin extrac, especially wherein the enzymatic teat fee pectin extract from the product of the (optionally) ‘enzymatic treated pectin comprising extract (Lethe proce tt obtained a c). With this process, advantageously in an embodiment polyphenol functionalized coffe pectin extract is produce, ‘whieh isa product that ean be sed for several applications, land which leads to @remisining product that has substan. \ially reduced content in polyphenols, and! may therefore be more easily reused or discanled as waste “The coffee pulp that is usod forthe process may directly be obiained from a plant, but may also have been subjected ta conservation process, The coffe pp used may also be joblained froma remote place (ike >10 km. or even >100 km ‘or even further, and after transportation be used as coffee pulp in the process of the invention. Before transportation the coffee pulp may optionally be treated for conservationsl purposes “The extraction per se, especially inchading an alkali snd acidic procedure, see also below, is also an aspect of the invention. Especially, the enzymatic treatment i appiod, whieh may be used to demethylate and/or cross-link The term "green coffee bean” is known in the art and especially refers tothe non-roasied coffee bean, The ceries that are used in de-pulping may be ina ripe or unripe state Also mixtures of unripe and ripe cheries may be applied ‘The properties ofthe polyphenol functionalized coffee pec tin extract may depend upon whether ripe andor unripe coffee beans are appli Pectin can be extracted from multiple sources, however pectins are mostly extracted from citrus peels and apple ppomace. As mentioned above, pets are chemically and/or US 9,896,572 B2 5 ‘exzymnaticaly modified to obtain desired gel structures Another source of pectin that has been accepted is pectin ‘extracted from the industal residues of sugar from beetroot (SBP). Physicochemical diflerences borwcon SBP and other ‘ype of conventional pectins include higher proportion of entra sugar side chains, a higher content of acetyl groups at O2 and O3 positions within the galacturonic backbone tnd a higher content af phenolic esters in the side chains particularly in the arabinose and galactose, and a higher ‘content of protinaceous materials bound to the side chains through covalent linkages. Unexpectely, colfee pectin shures some of the characteristics ininsie to SBP. the low molecular weight of the peetic molecules, the presence of ‘important amounts of galactose and arabinose inthe neutral side chain, and the presence of polyphenols among others. It is therefore theorized that eoflee pectin can be modified as SBP and yield high valve pectins with emulsifying charac- teristics. Also collee pectin ean be chemically modified as 7 ‘As can be derived from the above, in an embodiment the ‘method may include extrcting (irom the pectin comprising ‘griculturlby-prelact) pectin comprising exact, ‘wherein extraction is performed under acid conditions 10 provide the pectin comprising extract, followed by the enzymatic treatment, The additional extaction under alka- Tine conditions isa specie embodiment. As ean he derived from the above, also in an embodiment the method may include extrating (Irom the pectin comprising agricultural Dy-proiet) a pectin comprising extract, wherein extraction is performed under alkaline conditions, to provide the pectin comprising extract, followed by the enzymatic treaiment. The tlditional extraction under ace conditions is a specific embodiment. The acid extraction process especially pro- Vides a pectin that is useful for the food industry. The (daitonal dilute alkali extraction may assist in extracting peti that has low solubility in Water ‘As can he derived from the above, in an embodiment the ‘method may include extracting (Irom the peetin comprising ‘agricultural by-precuet) a pectin comprising. extract, ‘wherein extraction is performed under acid eonditions and alkaline conditions, t provide the pectin comprising extact whieh is a combination of the alkaline extraction product And acid extriction product, followed by the enzymatic treatment (of the combination of extracts). As will be discussed below, the acid extraction may be subsequent to the alkaline extraction or the alkaline extraction may be stibsequent to the acid extraction. The phrase “wherein extraction is performed under acid conditions and alkaline ‘conditions in general indicates that first an extraction is performed under acid or alkaline conditions and tht (sub- Sequenty) the remaining material from the acid or alkaline extraction is subjected to an alkaline or acid extraction, respectively. The extracts can be combined for further (enzymatic) processing and the remaining material ean be ‘used for other applications or discarded (sce elsewhere herein), Tina prefered embodiment, the acid coitions of the first extraction ae at a pH in the range of0.5-4, especially 1.53. Furer, de frst extraction may especially be performed at ‘temperature of at least 80°C. The alkaline conitions inthe second extraction are especially ata pH in the range of 7-14 seh as 8-14, even more especially 7-11, such a8 9-11, sueh fs expecially between 75105, such a 9,510.8. For the alkaline extraction, the pH is >7, especially 75-9. At larger PH, the pectin molecule may start getting hydrolyzed, Fur- ther, especially the alkaline extraction is performed at a lemperature not higher than 48° C. especially 35° C. ‘Further, in embodiments the one of more enzymes are selected from the group consisting of diphenol oxidoredne- tase, peroxidase, laccase, pecin-estorase, methylesterase, poly galacturanase, endo polyglucanase, and exo polyghi- cease. Alternatively or aitionaly, one oF more enzymes selected from the eroup consisting of pectin Ivase, poly Jacturonase (endo and exo), endo galaetansse, exo galt nase, rhamnogalacturomise nay be applied. Alternatively or additionally, especially arabinofuranosidase, arabinase, {eruloyl esterase, endo pectin methyl esterase, exo pectin ‘ethyl esterase, pectin esterase, lncease, peroxidase (espe- cially from horseradish) may be applied. For demethylation. ‘expecially enzymes like pectin methyl esterase (FC 3.1.11) ean be applied. For cross-linking, especially enzymes like US 9,896,572 B2 7 peroxidase (especially BC 1.10 or BC LI, such as eg. horseradish peroxidase 1.11.17) may be applied. Especially, the enzymatic treatment atleast involves ireatment of the ‘extraci(6) with an oxidoreductase. An oxidoreductase is an ‘enzyme that catalyzes the transier of electrons from one ‘molecule (reductant or cleetron donor) to another the mol- fecule (oxidant or electron acceptor). Best results ate ‘obtained with an oxidoreductase selected trom the (stb) ‘lasses EC 1.10 (oxidonedctases tat act on diphenols and related substances as donors) and EC 1.11 (oxidoreductases that act on peroxide as an acceptor (peroxidases). Espe- 7, especially 75-9. At larger pH, the pectin molecule may start geting hydrolyzed. Further, especially the alkaline extrsction is performed at s temperature not higher than 45° C. especially 35° C. Further, especially the cid conditions of the second extraction are ata pl in the range ofS, especially 1.5, In further embodiments, the second extraction s especially pefomed at a temperature of at Teast 80° C. The enzyme may be used in one of more ofthe following ‘instances: during the acid extraction, after the acid extrae- tion, during the alkane extraction alter the alkaline extac- tion, and during & stage when both extacts have been ‘combined. Of course, during one of more ofthese stages, an ‘enzyme may be applied. Especially, the enzyme, and ‘optional additive forthe enzyme such as HO. may depend upon the pH andior temperature. HO, may fr instance only be applicd when peroxidase is used, especially horse radish peroxidase, Lavease does not need HO, 10 generate the 0 o 8 polyphenol crosslinks, However, laccase in general only Substantially acts at pid between about 6.0 10 85. Horse radish peroxidase aets in general only substantially at pif higher between about 8.5 and 12.5. The extraction pH may thus eg, alko depends in tho enzyme used, though, if necessary, afer extraction the pH may also be altered to arrive ata pH suitable forthe chosen enzymes. Therefore, ia embodiments wherein (horse radish) peroxidase is applic the presence of HO. is desired and the pH range, during at least part ofthe process is especially fom 60 to 11 since this the range where the reduetase is more active. The ‘temperature in the alkali exircton is expecially not over 45° CC. Further, the optimum temperature for both Iaeease and peroxidase is in the range of 30-40° C. such as about 35°C. Enzymes may be added during any stage of the process, b fare of course at east available diring the eavymatic treat ‘meat ‘Ye in further embodiments, the one or more enzymes are expecially selected from the group consisting of diphenol oxidoreductase, peroxidase, nvease, — pectin-sterase, methyl-esteras poly galacturanase, endo polyglucanase, and exo polyglucanase. Alternatively or additionally, one oF more enzymes are selected from the group consisting of pectin lyase, polygalacturonase (endo and exo), endo galae- fanase, exo galactanase, rhamnogalacturonase may be applied. Alternatively or additionally, especialy arabino- furanosidase, arsbinase, feruloy! esterase, endo pectin ‘methyl esterase, exo pectin methyl esterase, pectin esterase, Jacease, peroxidase (especially from horseradish) may be applied. For demethylation, especially enzymes like pectn- fsterase con be applied. Por crost-linking. especially enzymes like peroxidases may he applied, As indicated hove, during the enzymatic treatment expecially atleast an oxidoreductase selected from the (subjelasses EC 1.10 and TEC 1.1 is applied. Note that che term “an enzyme” or “an oxidoredctases” and simile terms may also refer to a plurality of (ifferent) enzymes or a plurality of (ifferen) oxidoreductases, ee, respectively. As indicated herein, the fenzyimatie treatment may for instance be during an extac- tion stage or subsequent to an extrction stage, or multiple enzymatic restments may be appli. Further, also cock- tail of diferent enzymes may be added, Assuming a ox reductases, the amount of enzyme used will be in the range of about 0.5-10 mg, such as especially about L mg of pure enzyme (100% protein) per 100 ml and essuening aa esterase, the amount of enzyme used willbe in the range of about 1-10mg, especially about S mg of pure enzyme (100% protein) per 100 mi Fence, the enzymatic modification may be executed in a reactor, whore there san setual transfor. ‘mation of the matter. In one or more of the alkali and acid extraction there may be no (enzymatic) modification. There- Tore, these may be executed inn extraction unit. During the cenyimatie modification slep the enzyme(s) may especially transform the peetin to a cross-linked pectin andor may cleave groups attached to the pectin (macromolecules). ‘In a further spocitic embodiment, prior to the (first) extraction, the coe pp i subjected to a washing process, ‘wherein the washing process comprises mixing the coffee pulp with a solvent and subsequently removing atleast part fof the solvent, wherein the water content of the solvent is +280 wi, %, Especially, at least 50 wl, %, even more especialy at feast 80 wt. 9%, ofthe solvent consists of one or more liquids having a polarity lower than water (vide inf), Tn an embodiment, the extraction of polyphenol fintion- alized coffee pectin extract from the product of the option- ally enzymatic trated pectin comprising extract comprises ing at least part of the enzymatically treated material US 9,896,572 B2 9 With an extraction liquid and subsequently removing atleast part ofthe polyphenol funetionalized coffee pect, wherein the extraction liguid has a pH inthe range of 35-6, such as 4-6, Especially, the extmcton liquid comprises ethanol However, the exiziction liquid may also comprise other solvents, such as methanol, 2-propanol, acetone. The same typeof solvent may be used as used forthe washing process (Gee also helow). Funher, the extraction liquid may be ‘scidtiod. Further, the extraction Tiguid may comprise 9 ‘combination of two or more of (such) solvents. This extrac tion liquid may be used to precipitate the funetionalized pectin. By adding the solvent, pectin may precipitate cret- fing a gel which can be separated from the low molecular ‘weight compounds dissolved in the solvent ‘With the process ofthe invention, but optionally via other routes, @ polyphenol functionalized coffee pectin extract may be oblained. Hence, in 2 further aspect, the invention also provides a polyphenot functionalized pectin (per se), ‘especially a polyphenol functionalized coffe pectin. Hen ‘especially he invention provides « polyphenol factional ‘ned coaflee pectin obtainable by the process as deseribed herein. Especially the polyphenol functionalized cee pec- tin has a molar ratio of phenolic groups to the sum of arabinose pls galactose units between 20% to 60%, and has ‘4 molecular weight 290,000 Da. In the pectin (product) of the invention, the protein content may be inthe range of $-18 wt. % especially 8-15 wt, %, Further, the polyphenol ‘content may be in the range of 0.06-0.18 wt. %, espocially £0,09-0.15 wt. %.As knows in thea, the protein content ean be determined based on the Dumas methiod (SO 16634-1 2008); the polyphenol content can be determined based on the Folin-Cincalten (ISO 14502-1:2005) method with the Folin-Ciocalteu reagent (FCR) or Folin’s phenol reagent or Folin-Denis reagent, also called the Gallic Acid Equivalence method (GAF). This reagent (method) is especially designed for determining the phenol amount. Fspecaly, he invention Jurher provides polyphenol funetionalize! coffee postin (es described herein), having a molar weight =200,000 Da, sueh as especially 120,000 Da, such a6 in the range of 90,000-120,000 Da. The cof pectin may have a total ugar ‘content of rhaninose, arabinose, xylose, mannose, galactose, tlucose, galacturonic acd, relative othe total sugar content, Jn the range of 70.95 wt, °%, with a total galacturonic acid ‘content, relative to the oll sugar conten, ia the rine of 5S ‘0 80 wt. %. Further, the total glucose content, reatve to the total sugar cootent, may be inthe range of 3-15 wt. %. The ‘otal sugar content in the pectin (product) of the invention may be in the range of 40.80 wt. %, relative to the total ‘wojght of the product. The remaining part may inelude amongst others polyphenol and protein, Further, especially the Gal/UA ratio may be in the range of 0.1-0.2(galactose- uroaie acid weight ratio), In addition to the above, further (intermediate) process steps may he included, such as one oF more of precipitation, fikration, washing and resaspension. For instance, after recombination of the two extriets (or ater acid extraction, but) before an enzymatic treatment, (also) a precipitation, ‘ration, washing and resuspension may take place. The terms “upstream” and “downstream relate here to an ‘erangement of items oF features, of & sequence of stages, relative to the propagation of a process chain, wherein relative ta fist stage within a chain of process aetions oF process apparatus oF process stages, a second stage in the process chain closer to the beginning of the chain is Sopstream. and a third stage within the process chain ure away from the process beginning is “downstean 0 o “substantially s", will be under- stood by the person skilled in the art. The term “Substan- tially may also include embodiments with “entirely” “completely”, “all, te. Hence, in embodiments the adjee- tive substantially may’ also be removed. Where applicable, the term “substantially” may also relate (0 90% or higher such as 95% of higher, especially 9% or higher, even more 99.5% of higher, including 100% The term inchides also embodiments wherein the term “eomprises” means “consists of". The tenn “and/or” espe- cially relates to one of more ofthe items meationed before and after “andr”. For instance, phrase “item 1 andr ite 2° and similar phrases may relate to one or more of item ‘and item 2. The term “eomprising” may in an embodiment refer to “consisting of” but may in another embodiment sso refer to “containing at least the defined species and option- ally one or more other species” FFurtbermor, the terms fist, second, tht! and the Tike in the desription and in the claims, are uted for distinguishing between similar elements and not necessarily for deseribing ‘sequential or chronological order. It is to be understood that the terms so used are interchangeable under appropriate circumstances and that the embodiments of the invention described herein are capable of operation in other sequences than deseribed or illustrated herein, ‘The apparatus herein are amongst others deseribed during ‘operation. As will be clear tothe person skilled in the art, the invention is oot limited to methods of operation or devices in operation shouldbe noted thatthe above-mentioned embodiments illustate rather than limit the invention, and that those skilled in the art will be able to design many altemative embodiments without departing from the scope of the ‘appended claims. In the claims, any reference signs placed between parentheses shall not be construed as Himiting the claim. Use of the verb “to comprise” and its canjugations {des not exclude the presence of elements or steps other than those stated ina claim. The antcle “a” or “an” preceding an element does not exclude the presence ofa plurality of sueh elements. The invention may be implemented by means of hardware comprising sever distinct elements, and by ‘means of suitably programmed computer. Inthe device claim enumerating several means, several of these means ‘may be embodied by one and the same item of hardware The mere fact that certain measures are recited i mutually erent dependent claims does not indicate that a combi- pation of these measures cannot be used to advantage. ‘The invention further applies to an apparatus comprising ‘one oF more ofthe charocterizing features described in the Seseription andor shown in the attached drawings. The invention further pertains toa method or process comprising ‘ne of more of the characterising features described in the ‘description andor show in the attached drawings. The various aspects discussed in this patent can be combined in onder to provide additional advantages. Fur- thermore, some of the features can form the basis for one oF more divisional applications. BRIEE DESCRIPTION OF THE DRAWINGS Embodiments of the invention will now be deseribed, by \way of example only, wih reference to the accompanying schematic drawings in which corresponding reference sym- bols indicate corresponding pars, and in whieh: FIG. 1 schematically depicts an embodiment of the pro- ‘ess as described here US 9,896,572 B2 un FIGS. 24-26 schematically depiets coffee pectin (before and affer processing as deseribed in some embodiments herein) TIG. 3 shows a high performance size exclusion chrome ‘ography (HPSEC) for commercial pectin from cits pool and postin obtain with the herein deseribed process: FIG. 4 shows the presence of oligomers aller digestion of pectin with polygalacturonase from Aspergiius aculets ‘aller high performance ‘anion exchange chromatography (HPAEC) for commercial poctn from citrus peol and pectin ‘obtain with the herein deseribed process TG. Sa schematically shows a coffee bean: FIG. $4 shows a (Laboratory) scale process for fresh material; FIG, Se shows the oxidation of cathecol by PPO from coffee polp in time (in absorbance units, y-axis, and with time in hours on the x-axis). Data are obtained spectropho- ‘ometrically at 420nm. The sing © indicates the absorbance ‘change of the enzyme extract (indicated with square), minas the absorbance change of the substmte soliton (indicated ‘with triangle), without enzyme: FIG, 8d shows a low chart for large-scale preservation of ied coffee pulp: and TIG, Se shows a large seal extraction from preserved wet pulp (with mass balance). The schematic drawings herein ‘ae nol necessarily on scale, DETAILED DESCRIPTION OF THE PMBODIMENTS, FIG. 1 schematically depicts an embodinent of the pro ‘ess as described herein, Blended caller pu the collec Pilly may relate to one or more of the exocamp, ster mesocap (the polp isl, andthe mesocarp (ameiage or parchment), The ul (abo knowa ssiver skin) is not taken into account ater during he process, huling machines may be wed to remove the parchment layer) from wet processed ces Plping dry process coffe refers to removing the ‘otre deed isk the exoearp, mesoearp & endocsep of the dried cheies. Further, optionally polishing may tke lice: this an optional process in which any sir skin tat Femains on the heans afte pulping (optionally a polishing tuachine may be applied). Tae pulp is prefebly collected From the depulping mills soon as possible alter aenertion ‘ofthe pulp, peterbly i the st 24 hours, Would this aot be possible, itis prefered to use preservation steps 10 minimize pectin depadation de wo enzymatie activity. The preservation tps cam be but ae ot nite to: aeiteation Df the pulp toa pH below 4, alkaiiston toa pl above 9 lowering ofthe water activity 1 ae-0.5 or less at tempera: tures blow 60” C- inactivation of dae endogenous enzymes by solvent adltion, inetivation ofthe endogenous enzymes by boiling andor cooling (eyeles). Adon of one or more preservation agents solcted fom the group consisting of fodinm mets bislfite, ascorbic acid, elbyenediamintet Tascatie aid EDTA) may also be apt. Examples of (cher preservation sents ar sled rom the soup consisting of ascobie ai. cre acid, oxalic aed, sium metabislite, potassium bisulfite, sular Gioxie, lyeine, methionine and EDTA, especially one or more of sodium nmetzbisulie, potassium bisulfite, and EDTA. ones, in embodiments of the process, prior 1 the ceatation hut especialy afer washing) the pp is subs jected to a preservation process, wherein the preservation Process comprises one or more of () adding & preservation gent to the pulp and (i) dying the pup. As indicated ‘above, the prckervation agent may comprise one or more of o 12 ascorbic acid, etre acid, oxalic acid, sodium metabisulfive, potassium bisulfite, sulfur dioxide, glycine, methionine and EDTA, especially one or more of sodium metabsulfive, potassium bisulfite, and EDTA, especially onc or more of sodium metabisulite and potassium bisulfite, Further, iis preferred tha the pulp is milled (or macerated) toa suitable particle size (after preservation but) prior o any processing ‘ep or stage. suitable particle size i in the range between TO.and 40, such as eg. 18 mesh. The cofie pulp is indicated with reference 10, The wse of milled pulp my lead to a better exiaction dhan when sing unmilled pulp ‘The (optional) clean-up procedure, herein also indicated as washing process, indicated with reference 100, may comprise the treatment ofthe (blended) coffee pulp with a solvent of low polarity, it can be, but is nt limited to one or wore of acetic acid, acetone, acetonitrile, acetyl acetone, 2-aminoethanol, aniline, anisole, benzene, benzonitrile, ben: yl alcohol, I-butapol, 2-butano, i-butanol, 2-butanone, ‘Churyl alcohol, carbon disulfide, carbon tetrachoride,chlo- rubenzene, chloroform, eyelohexane, eyelobexanol, eyelo- hhexanone, d-n-butylphthalas, I. 1-dichloroothane, diethy!- ene glycol, I-Methony-2(2-methoxyethoxy)-cthane (diglyme). dimethonyethane (glyme), N.N-dimethy! aniline, dimethyl formamide (DMF), dimethyl phthalate, dimethyl sulfoxide (DMSO), dioxane, ethanol, eer, ethyl acetate, ethyl acetoacetate, chy benzoate, ethylene glyeo!, glycerin heptane, I-heptanol, hexane, I-hexanol, methanol, methyl ‘acetate, methy] t-butyl ether (MTBE), methylene chloride, TFoctanol, pentane, I-pentanol, 2-pentanol, +-pentanal, 2-pentanone, 3-pentanone, I-propanol, 2-peopanol, pyri- dine, tevahydrofuran (THE), toluene, suchas especially one for more of methanol, methanol water mixture, hexane toluene, ethylene glycol, ether, ethyl ether The solvent may ‘optionally be acidified. The pulp is mixed with the solvent, preferably ina counter current extractor. The hydrocinnamic ‘acids as well as the free polyphenols are dissolved. In this Stream there may also be &rih fraction of effeine which in Jater stage can be purified, and might bea sub product ofthe pectin extraction. The pricict obtained after the washing process is indicated with reference 110, Washing with 9 Solvent may remove fee polyphenols and caffeine trom the coffee pulp and it may also precipitate polysaccharides of higher degree of polymerization. Washing with slvent may remove as much free polyphenols a possible without removing te polysecharides that have polyphenols i their fimetional groups. Bottom line is solvent of lower po such as ethanol and propanol, precipitate the pectin with polyphenols attached while sohbilizing caffeine and poly- Phenols that sre not attached to the pectin sirutire. This allows to later use enzymes such as lacase to selectively ‘modify pectin with biphenolic groups without loosing 100 ‘much enzyme polymerizing the high amount of polyphe- sols. The washing liquid for the pulp may especially com- prise loss than 70 wt. % water, especially fess than 68 wt. % Water, such as less than $5 wt. %, of even lovver. The Jiquid(s) used forthe extractions) especially have a higher ‘ater content than the washing liquid (lor the pulp). For instance, the extraction liquids) may comprise more than 6S ‘Wt %, expecially more thaa 75 wi. %, even more especially at leat 80 wt. %, such as atleast 90 wr. % ike at least 9S ‘wt % water Further, the liquids) used forthe extaction(3) tespecially have a higher polarity than the washing liguid (for the pulp). In this way, feee polyphenols and caffeine may be removed fiom the pulp ty the washing liquid and. by extraction with a polar solvent (especially an acidified) polar solvent) pectin may be extrcted from the insoluble solids ofthe pulp, Further, a indicated above, the extraction US 9,896,572 B2 13 Tiguid(s) expecially has a piI<7 or a pl1>7. Optionally, the ‘extiction igus) may also comprise a (solved) salt (see ‘ako below). Tina first extraction stp or stage, indicated with reference 200, the biomass, especially the product obtained after the ‘washing process, may in an embodiment, be acidified to 9 pll-4 or lower, especially a plI=2 or lower, with a concen- trated acid such as but not limited to one or more of hydrochloric acid, nitrie acid, phosphoric acid, acetic acid. Further the mixture may be heated, sul as toa temperature ‘of atleast 70° C. The pH is preferably lowered before the heating step or slage eat be applied. The ratio of biomass to (extraction) liquid may especially be in the range of 0.25 1-1:025, especially 0.5:1-1:0.5, such as 1:1 (which means that foreach kilogram of fesh pulp one lite of solution is necessary for the extraction). It is preferably to use a high ‘concentrated buffer sofution to mix the biomass and then adjust the pH, Possible salts solutions are (but aot limited) ‘o one of more of sodium mono basi phosphate (NaHPO,), sodium nitrate, sodium acetate, and sodium chloride. Alter 2 natively or additionally, ammonium andor potassium salts may be applied. OF course, more than one salt may be applied. The concentration of the salts may mnge between 50-400 mM, such as especially 100 mit t 200 mM (for ‘each salt individually), Extraction may e.g. be executed in an ‘extrction vessel ora counter eurent extractor where liquids and solids are mixed together and mixed continuously. I is desirable that heating is performed as fast as possible, ‘Thorefore, pre heating of the extraction vessel may be advisable. The product obtained after the frst extraction step fr stage is indicated with reference 210. It is preferred that the fist separation step or stage indicated with reference 300, s performed on the hot mix (obtained inthe frst extraction step or stage. Itis desired t0 recover a much solution (ie. fate oF supernatant) as possible before continuing with the next step oF stage. ‘According tothe setup and magnitude ofthe stream different types of separation units ean he used. Examples ae frame separator plate sepamtor, a sieve (separtor) and a cen- tefuge (separator) The solid precipitating from a liquid is called a precipitate (residual product or fist residual prod ‘uct, oF When compacted by a centrifuge, a pellet. The liquid remaining above the solid is in either case called the ‘upernte oF supernatant, Also filtration with filter may be performed. The process of passing a mixture through a iter {s called fileation. The liquid produced after filtering, ia zener a suspension ofa solid in lguid, i called trate, ‘while the solid remaining in the iter is called retentate, residue, of fltand. The remaining liquid after the fst Separation, the supernatant or filtrate (here the fist extrac- tion product), goes to a reactor in which it may be mixed With the supernatant of a second separation step or stage. ‘The precipitate, retentate, esidue, sediment or Hlirand (st residual prot) must especially be composed of only solid matter as mich as possible. At this poiat the (remsining) biomass should be approximately $0% 0 75% of thestaring mass (dry weight. The products obtained after the fist separation step oF stage are indicated with references 310 and 320, Reference ‘310 refers © the product that is remaining after the first, ‘separation, such asa retenat, residue or iltrand, sediment, ‘te. This product 310 (fist rescoal product) is especially subjected to a second extraction 400, see also below. The (desir) product, indicated with reference 320, of the Separation action, i. a filrate or permeate or supernatant ‘etc (first extraction product) can be directly introduced in 3 first reactor 600 (or reaction staze), see also below. The 0 o 14 pectin comprising (extract after separatos ‘meat indicated with reference 320, is a liquid product (extraction liquid with extact) In a second extraction step oF stoge, indicated with reference 400, the first exiriction residual product 310 oF biomass may he mixed with alkali to extract the more ramified polysaccharides as well as more esterified pectin ‘hich comprised the coffee pulp. This may in an embod ‘ment be done in several steps or stages. First with a(a extraction) liquid, especially water, the biomass is diluted ‘until 50% ofthe total dilution is achieved, Thereafer, the pH may be adapted, eg. with concentrated alkali (Le. an alkali solution), toa value of especially a least 9, lke eg. 10. ln ‘an embodiment, aller adding the alkali, hydrogen peroxide ‘may be sed up to especialy a concentration of up to 7.5%, especialy up to 5% ofthe stating biomass. In the last stage of this extraction, the volume is completed with water Exttgetion may be executed in an extraction vessel, extraction vessel may be composed of a recipient” in adequate material, such 2s stainless sfel 320, 316 or alloys that prevents rust A recirculating pump may or may not be present depending of the operation if cantines or batch The extraction vessels must inchide a source of heat and mixing ‘mechanism. Mixing should be promoted to obiain higher rates of delignification and hydrolysis of the esterified ‘compounds attach to the pectins. The temperature of the extraction vessel is especially regulated to avoid breakdown ofthe biopolymer. suitable temperatre is inthe range of 35.65" C. The alkali concentration (of the concentrated alkali solution) is especially approximately 6 molar to 8 ‘molar. The alkali (Solution) ean he based ona solution of eg, fone or more of sodium hydroxide, potassiam hydroxide calcium carbonate, and ammonium acetate. The product (nixture) obtained alter the second extraction sep or stage 5s indicated with reference 410, Tn a socond separation step or stage the im is especially {0 separate the sols (om the produet (mixture) obtained after the second extraction), Le. the second (extraction) residual product, which ae mostly cellulose and lignin from the free polysaccharides that are in solution (second extrmc- ‘ion product, if necessary the pHT can be lowered to 7 of 8 before separation, lowering OF the pH! may be controlled {o (substantially) prevent gation ofthe peetns, which may lead to a loss of the biopolymers with the reientate. The second separation step or stage is indicated with reference 00. According to the setup and magnitude of the stream diferent types of separation units ean be used. Examples are frame separator, a plate separator, a sieve (Separator) and 8 centrifige (separator). The (desired) product of the (Sec- fond) separation action, ie. filtrate or permeate, or super ratant, ete. (12. the second extraction product), indicated ‘with reference §20, can (also) directly introduced in frst reactor 600. The second (extraction) residual product, not indicated, can be discarded "Note that in this schematically indicated process the second extniction stage 400 is downstream of the first reaction stage. Note however that the acid and alkaline extractions may also be performed in nother onder, ie. the fist reaction stage including an alkaline extraction and the second reaction stage including an acid extraction. The product obtained aftr alkaline and acid extraction (or acid an alkaline extraction), may also be relevant per se, How- fever higher quality pectins may be obtained sehen also the enzymatic processing as defined herein is applied. Alkaline extraction may optionally be omitted, acid extraction ha ever is especially desired US 9,896,572 B2 so indicated as reactor 600, both sacid-extrcted pectns aod f present alkali-etractd pectins may be mixed. Mixing may be done in ways known to the person skilled in the ar, like with an extruder ora stor. Hence, the frst reactor may especially include one or more ‘ofan extnider and a stirrer. Disc to the change of pH some pectns can gel. Fence, i is especially prefered 1 lower the PH (ofthe alkaline liquid) slowly ta point near neutrality, ‘especially in the range of 6-8 pli, preferably between pil 65-70. As indicated above, optionally directly afer acid ‘exirction, the enzymatic treatment may be executed. In uch instance, the pH of the separation product may be ‘increased to a pH in the range of 6-8 preferably between pFT 65-70. In the first reactor, especialy two types of enzymes, ‘oxidoreductase andr esterase, are applied separately o ‘combination. Oxidoreductase i added according to its activ ity, ie. the aecessary amount to react with the pectia polymer is added, Examples ofthe oxidoreductase are (but rot limited) wo: dipbenol oxidoreductase, peroxidase, and cease. Fsterase, especially pectin esterase, is added t0 control the degree of methylation and esterification of the postin. Example of the (esterase) enzymes are (bit not Jimited) to: pectinesterase, methyl-esterase, poly galacturs- nase, feruloyl esterase, arabinose, arabinofuranosidase and ‘endo and exo polyplvcanase (see also above) As indicated above, at least an oxidoreducatase may be applied, even ‘more especially in combination with an esterase (especially Pectinesterase (BC 3.1.11) In addition to the enzyme(s) ‘alka one or more further adkitives may be added, AS indicated above, the pH may he changed (if novessary) to ‘approximately neutral. Also other enzymes than defined herein, having the same functionality may be applied Te product obtained after processing inthe first reactor, fr aller processing inthis reaction stage, i indicated with reference 610. This produet may be subjected to a next reaction stage, which is indicated with reference 700. Herein, rferences 600 and 700 may’ (also) reer to different reactors, respectively. However, these references may also refer to reaction stages, which may in an embodiment be Performed consceutvely indifferent reactors whereas heat Source, pH control and thermostat are present or in the same reactor. Reference 700 may also refer herein to a second reactor (or vessel In a second rewotor (indicated with reference 700), a solvent, such as ethanol, methanol, 2-propanol, acetone ‘especially an acidified solvent, such as acidified ethanol (like ethanol 41% Acetie acid anhydrous) is added, espe- ally in the ratio of 121-1210, such as 1.2:16, Tike 1-4 extraction solution fo (acidified) solvent, such a6 ethanol “The function of the solvent ix expecially to change the polarity ofthe solution so the pectin will precipitate creating ‘ge which ean be separated om the low molecular weight ‘compounds dissolved in the solvent. The mix is left for ‘coagulation ofthe pectins and precipitation. I is preferred thatthe pH in this stage is lower than 6; however is not ‘advisable to have a pl lower than 3. I ngvessary, the solvent (or extraction liquid) preferably has a concentration of alcohol of 70% of higher. reactor is or comprises also decanter. In this way, the upper Jayer ofthe solution can be disposed leading to much smaller volume forthe lst separation step or stage. ‘Ina third separation step or stage indicated with reference ‘800, the unrefined pectin, indicated with reference 710, is, separated from the solvent(s), such as ethanol and coher solvents used during the process. Due to the colloidal charaeteristis ofthe peetns, in an embodiment a 0 o 16 ceatrifage may be applied for separating the pectns from the solvent(s). The product thus obtained (here the reentae, fikrand or sedimentation, ete.) is indicated with reference 510, which comprises the polyphenol functionalized (coffee) pect, The product for this reaction stage (or this reactor) is Indicated with reference 810, and can be indicated as the third extraction product (hich is a solid material). Avain, reference 800 may also refer to a further reactor, thi reactor which must he constricted in resistant material such as stainless stool 316, and 320, fire proof and suitable to ‘Work with volatile solvents. However, references 700 and ‘800 may’ also refer to a reactor including a decanter ‘fdesired, the pectins this obtained can further be refined to met specification in differen industries, For instance, higher molecular weight pectins can be obtained by furher crosslinking with (purified) enzymes andor gelling pectins ‘ean be de-esterified to met differen types of application in the food and beverage industries. Furhermore, pectin can be modified with arabinase and arabinofuranosidase fo obtain specific emulsification properties. "A very schematic drawing ofa pectin 20 from coffee pulp is indicated in FIG, 2a, MG refers to methyl group: AG refers to acetyl group, GUA refers to galacturonic seid, Gal refers to galactose, RHA refers to rhamnose, and Ara refers to arabinose, Reference 21 refers to the homogalacturonan region, reference 22 refers toa the shamanogalacturoan 1 region, and reference 23 refers to the neutral side chain region of pectns. FIG, 26 very schematically depiets a pectin obtainable with the process of the invention, wherein the pectins are crosslinked via eros-nk(s) CL. Reference Fer refers t0 ferulic acid (a phenol that is the basis of the cross-links, together withthe arabinose units), of which of each pectin, via the arabinose units the pectins may’ be eross-linked With the aid of fenulic eid. The enzyme oxidoreductases, sch as Jcease and/or horse radish peroxidase, may generate eross- Tinks in the fomn of polyphenols, such a diphenols or even polyphenols having more than fo phenol groups. EXPERIMENTAL Example 1 Extraction and modification of pectin from coffee pulp Laboratory scale procedure: Fresh pulp was obtained directly from farm in Colombia in the beginning of January 2012. The cherries were in ‘optimum ripe state tobe separated from the bean in the wet ‘ill, Aer the reollection the skin and pulp (exocarp and ‘meso carp) were separated with # manual. The pulp, skin (pulp) are blended and froeze dried for transport to The Netherlands. 1g of freeze dried material is washed 3 times ‘with acidified elhanol 80% and centrifuge at 3000 rp for $ ‘min in each step, the solids are then suspendad in water, the pHis adjusted 0 2.0 with hydrochloric aeidand fil t $0 ml Yolume. The suspension is shaken ina water bath st 70° ¢ Tor’ h The suspension is then ceatefnged at 3000 RPM for 10 min and the aqueous phase is separate from the solids, The solids are then neutralized and Sodium hydroxide is added to adjust pH at 10 and a final volume of $0 ml. The solids are suspended and shaken for [at room temperature Aer the alla extraction the suspension is centrifuged at 3000 RPM for 10 minutes andl the aqueous phase is pooled ‘with the acid solution, The pH is adjusted to 60 with diluted alkali or aid. The spent material is then dried for furher Analysis, The liquid obtained for extraction has a brownish colour. 3 ml of Fydrogen peroxide is (30%) is added to the US 9,896,572 B2 17 Fiquor and) $00 ul of horse radish peroxidase solution (5 ig/ml) is added and the solution is stined for 24 hours at room temperature. After the incubation 4 volumes of bso- Jute than are added and the pectin is lel to precipitate for 2 hour at 4 C. After precipitation the suspension is Hltered through a Whatman #3 filter paper wit the aid of Buchnee funnel. The retenate solide are washed with 100 ml of ‘acetone andl dried at 0° C. for 12 h with high convection, ‘The resting film is then milled in ball mill an stoned foe further analysis, Example 2 ‘The following procedure for obsaining soluble polysoe=

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