BR102020026250A2 - CITRIC ACID PRODUCTION PROCESS FROM WASTE PRODUCTS FROM THE SUGAR BEVERAGE INDUSTRY - Google Patents
CITRIC ACID PRODUCTION PROCESS FROM WASTE PRODUCTS FROM THE SUGAR BEVERAGE INDUSTRY Download PDFInfo
- Publication number
- BR102020026250A2 BR102020026250A2 BR102020026250-5A BR102020026250A BR102020026250A2 BR 102020026250 A2 BR102020026250 A2 BR 102020026250A2 BR 102020026250 A BR102020026250 A BR 102020026250A BR 102020026250 A2 BR102020026250 A2 BR 102020026250A2
- Authority
- BR
- Brazil
- Prior art keywords
- citric acid
- fermentation
- production
- drinks
- beverage industry
- Prior art date
Links
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 title claims abstract description 132
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 30
- 235000013361 beverage Nutrition 0.000 title claims abstract description 24
- 235000000346 sugar Nutrition 0.000 title claims abstract description 17
- 239000002699 waste material Substances 0.000 title abstract description 13
- 238000000855 fermentation Methods 0.000 claims abstract description 25
- 230000004151 fermentation Effects 0.000 claims abstract description 25
- 238000000034 method Methods 0.000 claims abstract description 23
- 150000001720 carbohydrates Chemical class 0.000 claims abstract description 9
- 235000014633 carbohydrates Nutrition 0.000 claims abstract description 9
- 241000228245 Aspergillus niger Species 0.000 claims description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 14
- 239000001963 growth medium Substances 0.000 claims description 12
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 9
- 150000008163 sugars Chemical class 0.000 claims description 9
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 6
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 6
- 235000014214 soft drink Nutrition 0.000 claims description 6
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 claims description 4
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 claims description 4
- 235000015203 fruit juice Nutrition 0.000 claims description 4
- 239000002054 inoculum Substances 0.000 claims description 4
- 239000011777 magnesium Substances 0.000 claims description 4
- 229910052749 magnesium Inorganic materials 0.000 claims description 4
- 238000012545 processing Methods 0.000 claims description 4
- 239000011701 zinc Substances 0.000 claims description 4
- 229910052725 zinc Inorganic materials 0.000 claims description 4
- 229910019142 PO4 Inorganic materials 0.000 claims description 3
- 235000015897 energy drink Nutrition 0.000 claims description 3
- 235000020400 fruit nectar Nutrition 0.000 claims description 3
- 229910052742 iron Inorganic materials 0.000 claims description 3
- 229910052757 nitrogen Inorganic materials 0.000 claims description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 3
- 239000010452 phosphate Substances 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 235000011496 sports drink Nutrition 0.000 claims description 3
- 238000013019 agitation Methods 0.000 claims description 2
- 238000011081 inoculation Methods 0.000 claims description 2
- 230000009469 supplementation Effects 0.000 claims description 2
- 241000228212 Aspergillus Species 0.000 claims 1
- 241000228257 Aspergillus sp. Species 0.000 claims 1
- 239000000758 substrate Substances 0.000 abstract description 14
- 241000233866 Fungi Species 0.000 abstract description 7
- 235000013305 food Nutrition 0.000 abstract description 7
- 239000000203 mixture Substances 0.000 abstract description 7
- 238000002203 pretreatment Methods 0.000 abstract description 7
- 235000019527 sweetened beverage Nutrition 0.000 abstract description 4
- 239000011785 micronutrient Substances 0.000 abstract 1
- 235000013369 micronutrients Nutrition 0.000 abstract 1
- 239000002994 raw material Substances 0.000 description 7
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 235000013379 molasses Nutrition 0.000 description 6
- 238000011282 treatment Methods 0.000 description 6
- 229910021645 metal ion Inorganic materials 0.000 description 5
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 239000002351 wastewater Substances 0.000 description 4
- 235000016068 Berberis vulgaris Nutrition 0.000 description 3
- 241000335053 Beta vulgaris Species 0.000 description 3
- 229930091371 Fructose Natural products 0.000 description 3
- 239000005715 Fructose Substances 0.000 description 3
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 3
- 229920002472 Starch Polymers 0.000 description 3
- 229930006000 Sucrose Natural products 0.000 description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 3
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 239000008121 dextrose Substances 0.000 description 3
- 230000002255 enzymatic effect Effects 0.000 description 3
- 239000000796 flavoring agent Substances 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 239000003755 preservative agent Substances 0.000 description 3
- 235000019698 starch Nutrition 0.000 description 3
- 239000008107 starch Substances 0.000 description 3
- 239000005720 sucrose Substances 0.000 description 3
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 235000008733 Citrus aurantifolia Nutrition 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 2
- 235000011941 Tilia x europaea Nutrition 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 230000001476 alcoholic effect Effects 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 2
- RYYVLZVUVIJVGH-UHFFFAOYSA-N caffeine Chemical compound CN1C(=O)N(C)C(=O)C2=C1N=CN2C RYYVLZVUVIJVGH-UHFFFAOYSA-N 0.000 description 2
- 239000011575 calcium Substances 0.000 description 2
- 229910052791 calcium Inorganic materials 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 229910002092 carbon dioxide Inorganic materials 0.000 description 2
- 235000021310 complex sugar Nutrition 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 235000019634 flavors Nutrition 0.000 description 2
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 2
- 235000021472 generally recognized as safe Nutrition 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 230000002335 preservative effect Effects 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- XOAAWQZATWQOTB-UHFFFAOYSA-N taurine Chemical compound NCCS(O)(=O)=O XOAAWQZATWQOTB-UHFFFAOYSA-N 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 241001513093 Aspergillus awamori Species 0.000 description 1
- 241000459887 Aspergillus luchuensis Species 0.000 description 1
- 240000006439 Aspergillus oryzae Species 0.000 description 1
- 235000002247 Aspergillus oryzae Nutrition 0.000 description 1
- 241000445051 Aspergillus welwitschiae Species 0.000 description 1
- 235000005979 Citrus limon Nutrition 0.000 description 1
- 244000131522 Citrus pyriformis Species 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- UYUXSRADSPPKRZ-UHFFFAOYSA-N D-glucuronic acid gamma-lactone Natural products O=CC(O)C1OC(=O)C(O)C1O UYUXSRADSPPKRZ-UHFFFAOYSA-N 0.000 description 1
- UYUXSRADSPPKRZ-SKNVOMKLSA-N D-glucurono-6,3-lactone Chemical compound O=C[C@H](O)[C@H]1OC(=O)[C@@H](O)[C@H]1O UYUXSRADSPPKRZ-SKNVOMKLSA-N 0.000 description 1
- 239000004267 EU approved acidity regulator Substances 0.000 description 1
- 108010073178 Glucan 1,4-alpha-Glucosidase Proteins 0.000 description 1
- 102100022624 Glucoamylase Human genes 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- LPHGQDQBBGAPDZ-UHFFFAOYSA-N Isocaffeine Natural products CN1C(=O)N(C)C(=O)C2=C1N(C)C=N2 LPHGQDQBBGAPDZ-UHFFFAOYSA-N 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- 239000005913 Maltodextrin Substances 0.000 description 1
- 229920002774 Maltodextrin Polymers 0.000 description 1
- 241000228143 Penicillium Species 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 240000000111 Saccharum officinarum Species 0.000 description 1
- 235000007201 Saccharum officinarum Nutrition 0.000 description 1
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000002535 acidifier Substances 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 239000003957 anion exchange resin Substances 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127090 anticoagulant agent Drugs 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 238000013452 biotechnological production Methods 0.000 description 1
- 229960001948 caffeine Drugs 0.000 description 1
- VJEONQKOZGKCAK-UHFFFAOYSA-N caffeine Natural products CN1C(=O)N(C)C(=O)C2=C1C=CN2C VJEONQKOZGKCAK-UHFFFAOYSA-N 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 239000003729 cation exchange resin Substances 0.000 description 1
- 229940023913 cation exchange resins Drugs 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 235000020971 citrus fruits Nutrition 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000010960 commercial process Methods 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 235000011850 desserts Nutrition 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 238000001784 detoxification Methods 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000012262 fermentative production Methods 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 229950002441 glucurolactone Drugs 0.000 description 1
- 150000004679 hydroxides Chemical class 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000000411 inducer Substances 0.000 description 1
- 238000011090 industrial biotechnology method and process Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 235000015110 jellies Nutrition 0.000 description 1
- 239000012978 lignocellulosic material Substances 0.000 description 1
- 239000004571 lime Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 239000006193 liquid solution Substances 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 238000003760 magnetic stirring Methods 0.000 description 1
- 229940035034 maltodextrin Drugs 0.000 description 1
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 description 1
- 230000037353 metabolic pathway Effects 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 238000006213 oxygenation reaction Methods 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 239000003973 paint Substances 0.000 description 1
- 235000019629 palatability Nutrition 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000005057 refrigeration Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 235000021309 simple sugar Nutrition 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000010907 stover Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 229960003080 taurine Drugs 0.000 description 1
- 235000014101 wine Nutrition 0.000 description 1
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 1
- 229910000368 zinc sulfate Inorganic materials 0.000 description 1
- 239000011686 zinc sulphate Substances 0.000 description 1
- 235000009529 zinc sulphate Nutrition 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
PROCESSO DE PRODUÇÃO DE ÁCIDO CÍTRICO A PARTIR DE DESCARTES DA INDÚSTRIA DE BEBIDAS AÇUCARADAS. A presente invenção refere-se a um processo de produção de ácido cítrico por fermentação submersa com fungos filamentosos utilizando descartes de bebidas açucaradas como substrato. O processo estabelecido pela presente invenção explora a composição rica em carboidratos e com baixas concentrações de micronutrientes dos descartes da indústria de bebidas açucaradas para utilização como substrato em fermentação para produção de ácido cítrico. A composição dessas bebidas permite seu uso como substrato em fermentação por fungos filamentosos para produção de ácido cítrico sem a necessidade de pré-tratamento. A invenção oferece uma alternativa economicamente e ambientalmente interessante para a indústria de bebidas açucaradas pela valorização dos produtos descartados, uma vez que além de reduzir a alta carga orgânica dos mesmos, há a produção de ácido cítrico, um produto de interesse comercial para as próprias indústrias de bebidas, indústrias de alimentos, farmacêutica e outras.PRODUCTION PROCESS OF CITRIC ACID FROM WASTE PRODUCTS FROM THE SUGAR BEVERAGE INDUSTRY. The present invention relates to a process for producing citric acid by submerged fermentation with filamentous fungi using sugary beverage waste as a substrate. The process established by the present invention exploits the composition rich in carbohydrates and with low concentrations of micronutrients of the sugary beverage industry waste for use as a substrate in fermentation for the production of citric acid. The composition of these beverages allows their use as a substrate in fermentation by filamentous fungi to produce citric acid without the need for pre-treatment. The invention offers an economically and environmentally interesting alternative for the sugar-sweetened beverage industry by valuing discarded products, since in addition to reducing their high organic load, there is the production of citric acid, a product of commercial interest to the industries themselves. beverage, food, pharmaceutical and other industries.
Description
[001] . A presente invenção refere-se a um processo de produção de ácido cítrico por fermentação submersa por fungos filamentosos utilizando descartes provenientes da indústria de bebidas açucaradas.[001] . The present invention relates to a process for producing citric acid by submerged fermentation by filamentous fungi using waste from the sugary beverage industry.
[002] . Ácido cítrico é um dos ácidos orgânicos mais utilizados industrialmente, especialmente na indústria de alimentos e farmacêutica em função do seu status de GRAS (geralmente reconhecido como seguro). O ácido cítrico é utilizado como conservante para minimização da deterioração oxidativa da cor e sabor de alimentos e bebidas, podendo também ser utilizado como agente emulsificante e aromatizante ácido. Geleias, sucos, refrigerantes, sobremesas, vinhos, produtos enlatados e em conserva são alguns exemplos de alimentos que podem ser adicionados de ácido cítrico em sua formulação como conservante e acidificante. Na indústria farmacêutica ácido cítrico é empregado para melhoramento da palatabilidade, mascarar sabores indesejáveis, como componente de sistemas efervescentes e como agente anticoagulante. Na indústria química, o ácido cítrico é utilizado em cosméticos, embalagens, produtos de limpeza, tintas, dentre outros (CIRIMINNA, R. et al. Citric acid: Emerging applications of key biotechnology industrial product. Chemistry Central Journal, v. 11, n. 1, p. 1–9, 2017.; DHILLON, G. S. et al. Recent Advances in Citric Acid Bioproduction and Recovery. Food and Bioprocess Technology, v. 4, n. 4, p. 505–529, 2011; SOCCOL, C. R. et al. New perspectives for citric acid production and application. Food Technology and Biotechnology, v. 44, n. 2, p. 141–149, 2006; VANDENBERGHE, L. P. S. et al. Production and Application of Citric Acid. In: Current Developments in Biotechnology and Bioengineering: Production, Isolation and Purification of Industrial Products. [s.l.] Elsevier Inc., 2016. p. 557–575).[002] . Citric acid is one of the most used organic acids industrially, especially in the food and pharmaceutical industry due to its GRAS (generally recognized as safe) status. Citric acid is used as a preservative to minimize the oxidative deterioration of the color and flavor of foods and beverages, and it can also be used as an emulsifying and acidic flavoring agent. Jellies, juices, soft drinks, desserts, wines, canned and preserved products are some examples of foods that can be added with citric acid in their formulation as a preservative and acidifier. In the pharmaceutical industry, citric acid is used to improve palatability, mask undesirable flavors, as a component of effervescent systems and as an anticoagulant agent. In the chemical industry, citric acid is used in cosmetics, packaging, cleaning products, paints, among others (CIRIMINNA, R. et al. Citric acid: Emerging applications of key industrial biotechnology product. Chemistry Central Journal, v. 11, n. 1, p. 1–9, 2017.; DHILLON, G. S. et al. Recent Advances in Citric Acid Bioproduction and Recovery. Food and Bioprocess Technology, v. 4, n. 4, p. 505–529, 2011; SOCCOL, C. R. et al. New perspectives for citric acid production and application. Food Technology and Biotechnology, v. 44, n. 2, p. 141–149, 2006; VANDENBERGHE, L. P. S. et al. Production and Application of Citric Acid. In: Current Developments in Biotechnology and Bioengineering: Production, Isolation and Purification of Industrial Products. [s.l.] Elsevier Inc., 2016. p. 557–575).
[003] . A produção comercial de ácido cítrico teve início por volta de 1826 com a extração a partir de frutas cítricas. Os primeiros processos comerciais, implantados na Inglaterra e Itália, envolviam a extração de ácido cítrico de suco de limão ou lima (GB423668A, GB136979A). Novos processos foram comercialmente implantados a partir da descoberta da capacidade de alguns microrganismos em produzir ácido cítrico em condições específicas. A produção biológica de ácido cítrico, especialmente empregando cepas do fungo filamentoso Aspergillus niger, se mostrou uma alternativa mais viável (US1066358A). Inicialmente, a produção biológica empregou o método de fermentação líquida em superfície. Atualmente, a maior parte do ácido cítrico é produzida por fermentação submersa pelo fungo Aspergillus niger. A produção de ácido cítrico via fermentação submersa permite a utilização de matérias primas amplamente disponíveis e de baixo custo, tais como melaço de cana e beterraba, xaropes, hidrolisados de amido e outros (IKRAM-UL, H. et al. Citric acid production by selected mutants of Aspergillus niger from cane molasses. Bioresource Technology, v. 93, n. 2, p. 125–130, 1 jun. 2004; SHOW, P. L. et al. Overview of citric acid production from Aspergillus niger. Frontiers in Life Science, v. 8, n. 3, p. 271–283, 2015; WANG, B. et al. High-efficient production of citric acid by Aspergillus niger from high concentration of substrate based on the staged-addition glucoamylase strategy. Bioprocess and Biosystems Engineering, v. 40, n. 6, p. 891–899, 7 abr. 2017). Os documentos supracitados reportam diversos fatores que influenciam a produção de ácido cítrico por fungos como A. niger em diferentes substratos e matérias-primas. Porém, a composição complexa dos descartes da indústria de bebidas açucaradas diferencia esses substratos dos demais reportados na literatura. Relação carbono/nitrogênio/fosfato, balanço de íons, presença de reguladores de acidez, corantes, edulcorantes, antioxidantes, mistura de carboidratos, maltodextrina, cafeína, taurina, glucoronolactona, aromatizantes naturais e artificiais, vitaminas, conservadores, dióxido de carbono são fatores presentes nos descartes que afetam a produção de ácido cítrico. Tais compostos podem ter efeitos inibidores no crescimento de A. niger ou promover desvios na rota metabólica do fungo. Desse modo, é necessário desenvolvimento tecnológico específico para esse tipo de substrato.[003] . The commercial production of citric acid began around 1826 with the extraction from citrus fruits. The first commercial processes, implemented in England and Italy, involved the extraction of citric acid from lemon or lime juice (GB423668A, GB136979A). New processes were commercially implemented from the discovery of the ability of some microorganisms to produce citric acid under specific conditions. Biological production of citric acid, especially using strains of the filamentous fungus Aspergillus niger, proved to be a more viable alternative (US1066358A). Initially, organic production employed the liquid surface fermentation method. Currently, most citric acid is produced by submerged fermentation by the fungus Aspergillus niger. The production of citric acid via submerged fermentation allows the use of widely available and low-cost raw materials, such as cane and beet molasses, syrups, starch hydrolysates and others (IKRAM-UL, H. et al. Citric acid production by selected mutants of Aspergillus niger from cane molasses. Bioresource Technology, v. 93, n. 2, pp. 125–130, 1 June 2004; SHOW, P. L. et al. Overview of citric acid production from Aspergillus niger. Frontiers in Life Science. , v. 8, n. 3, p. 271–283, 2015; WANG, B. et al. High-efficient production of citric acid by Aspergillus niger from high concentration of substrate based on the staged-addition glucoamylase strategy. Bioprocess and Biosystems Engineering, v. 40, no. 6, p. 891–899, 7 Apr. 2017). The aforementioned documents report several factors that influence the production of citric acid by fungi such as A. niger in different substrates and raw materials. However, the complex composition of waste from the sugary beverage industry differentiates these substrates from others reported in the literature. Carbon/nitrogen/phosphate ratio, ion balance, presence of acidity regulators, dyes, sweeteners, antioxidants, carbohydrate mixture, maltodextrin, caffeine, taurine, glucuronolactone, natural and artificial flavorings, vitamins, preservatives, carbon dioxide are present factors in wastes that affect citric acid production. Such compounds may have inhibitory effects on the growth of A. niger or promote deviations in the fungal metabolic pathway. Thus, specific technological development is necessary for this type of substrate.
[004] . A maioria das matérias primas atualmente empregadas necessitam de pré-tratamento. No caso do melaço de cana e beterraba, há necessidade de diversos tratamentos para remoção de íons metálicos e outros inibidores que prejudicam o desempenho do processo fermentativo. Matérias-primas constituídas de açúcares complexos necessitam de pré-tratamento para disponibilização de açúcares simples capazes de serem fermentados pelos microrganismos (ZHOU, P. P.; MENG, J.; BAO, J. Fermentative production of high titer citric acid from corn stover feedstock after dry dilute acid pretreatment and biodetoxification. Bioresource Technology, v. 224, p. 563–572, 1 jan. 2017; RIVAS, B. et al. Submerged citric acid fermentation on orange peel autohydrolysate. Journal of Agricultural and Food Chemistry, v. 56, n. 7, p. 2380–2387, 9 abr. 2008).[004] . Most of the raw materials currently used need pre-treatment. In the case of sugarcane and beet molasses, there is a need for several treatments to remove metal ions and other inhibitors that impair the performance of the fermentation process. Raw materials consisting of complex sugars need pre-treatment to provide simple sugars capable of being fermented by microorganisms (ZHOU, P. P.; MENG, J.; BAO, J. Fermentative production of high titer citric acid from corn stover feedstock after dry dilute acid pretreatment and biodetoxification. Bioresource Technology, v. 224, p. 563–572, 1 Jan. 2017; RIVAS, B. et al. Submerged citric acid fermentation on orange peel autohydrolysate. Journal of Agricultural and Food Chemistry, v. 56 , No. 7, p. 2380–2387, 9 Apr. 2008).
[005] . O documento CN104805136A descreve um método para a produção de ácido cítrico por fungo filamentoso utilizando como substrato material lignocelulósico. Mais especificamente, o documento se refere a um processo que envolve o cultivo de Aspergillus niger em hidrolisado enzimático de matéria prima celulósica ou por meio de sacarificação enzimática do substrato simultaneamente à fermentação. O processo de obtenção do hidrolisado enzimático envolve as etapas de pré-tratamento com ácido sob pressão e desintoxicação.[005] . Document CN104805136A describes a method for the production of citric acid by filamentous fungus using lignocellulosic material as a substrate. More specifically, the document relates to a process involving the cultivation of Aspergillus niger in enzymatic hydrolyzate of cellulosic raw material or by enzymatic saccharification of the substrate simultaneously with fermentation. The process of obtaining the enzymatic hydrolyzate involves the steps of pre-treatment with acid under pressure and detoxification.
[006] . CN101636500A descreve um método para produção de ácido cítrico a partir de glicerol. O processo descreve a utilização de glicerol em combinação com uma ou mais fontes de carbono, tais como melaço, glicose, frutose, sacarose ou amido hidrolisado por Aspergillus niger para produção de ácido cítrico pelo método de fermentação em superfície.[006] . CN101636500A describes a method for producing citric acid from glycerol. The process describes the use of glycerol in combination with one or more carbon sources, such as molasses, glucose, fructose, sucrose or starch hydrolyzed by Aspergillus niger to produce citric acid by the surface fermentation method.
[007] . A patente GB799752A descreve a produção de ácido cítrico por Aspergillus niger utilizando fontes impuras de carboidratos pelo método de fermentação submersa. Mais especificamente, o documento descreve a utilização de solução aquosa de melaço invertido, hidrolisado de amido de milho ou melaço de beterraba após tratamento para remoção de impurezas, principalmente íons metálicos. O pré-tratamento consiste na adição de cal ou hidróxidos insolúveis à solução aquosa rica em carboidratos (10 – 15% (m/v), seguido pelo tratamento com resinas de troca aniônica e/ou catiônica.[007] . GB799752A describes the production of citric acid by Aspergillus niger using impure sources of carbohydrates by the submerged fermentation method. More specifically, the document describes the use of an aqueous solution of inverted molasses, corn starch hydrolyzate or beet molasses after treatment to remove impurities, mainly metal ions. The pre-treatment consists of the addition of lime or insoluble hydroxides to the aqueous solution rich in carbohydrates (10 – 15% (m/v), followed by treatment with anion and/or cation exchange resins.
[008] . Na indústria de bebidas, uma considerável quantidade de bebidas é descartada ao longo da cadeia produtiva e comercial. Produtos são rejeitados durante o processamento por problemas relacionados ao padrão de qualidade, além disso, há uma parcela de produtos que retornam do mercado à indústria por problemas como perda de CO2 ou por ter expirado o prazo de validade (COMELLI, R. N. et al. Treatment of High-Strength Wastewater from the Sugar-Sweetened Beverage Industry by an Alcoholic Fermentation Process. Industrial and Engineering Chemistry Research, v. 54, n. 31, p. 7687–7693, 2015; ISLA, M. A.; COMELLI, R. N.; SELUY, L. G. Wastewater from the soft drinks industry as a source for bioethanol production. Bioresource Technology, v. 136, p. 140–147, 2013).[008] . In the beverage industry, a considerable amount of beverages is discarded along the production and commercial chain. Products are rejected during processing due to problems related to the quality standard, in addition, there is a portion of products that return from the market to the industry due to problems such as loss of CO2 or having expired (COMELLI, R. N. et al. Treatment). of High-Strength Wastewater from the Sugar-Sweetened Beverage Industry by an Alcoholic Fermentation Process. Industrial and Engineering Chemistry Research, v. 54, n. 31, p. 7687–7693, 2015; ISLA, M. A.; COMELLI, R. N.; SELUY, L.G. Wastewater from the soft drinks industry as a source for bioethanol production. Bioresource Technology, v. 136, p. 140–147, 2013).
[009] . No caso da indústria de bebidas açucaradas, os efluentes são ricos em carboidratos, o que requer tratamentos para diminuição da carga orgânica antes do descarte no meio ambiente. As bebidas descartadas durante o processamento ou após expirar o prazo de validade podem atingir até 135000 mg O2/L em níveis de demanda química de oxigênio (DQO). As bebidas açucaradas apresentam elevado teor de carboidratos (6 a 18% m/v), principalmente açúcares facilmente fermentáveis, tais como glicose, frutose, sacarose e maltose. Além disso, proteínas, cálcio, potássio, sódio, magnésio, zinco e vitaminas, dentre outros compostos, podem estar presentes. As bebidas açucaradas geralmente apresentam pH ácido variando de 2,5 a 5,0. Essa faixa é favorável à produção de ácido cítrico (COMELLI, R. N. et al. Treatment of High-Strength Wastewater from the Sugar-Sweetened Beverage Industry by an Alcoholic Fermentation Process. Industrial and Engineering Chemistry Research, v. 54, n. 31, p. 7687–7693, 2015; ISLA, M. A.; COMELLI, R. N.; SELUY, L. G. Wastewater from the soft drinks industry as a source for bioethanol production. Bioresource Technology, v. 136, p. 140–147, 2013; MAX, B. et al. Biotechnological production of citric acid. Brazilian Journal of Microbiology, v. 41, n. 4, p. 862–875, 2010)[009] . In the case of the sugary beverage industry, the effluents are rich in carbohydrates, which requires treatments to reduce the organic load before being discharged into the environment. Beverages discarded during processing or after the expiration date can reach up to 135000 mg O2/L in chemical oxygen demand (COD) levels. Sugary drinks have a high content of carbohydrates (6 to 18% w/v), mainly easily fermentable sugars, such as glucose, fructose, sucrose and maltose. In addition, proteins, calcium, potassium, sodium, magnesium, zinc and vitamins, among other compounds, may be present. Sugary drinks usually have an acidic pH ranging from 2.5 to 5.0. This range is favorable for citric acid production (COMELLI, R. N. et al. Treatment of High-Strength Wastewater from the Sugar-Sweetened Beverage Industry by an Alcoholic Fermentation Process. Industrial and Engineering Chemistry Research, v. 54, n. 31, p. . 7687–7693, 2015; ISLA, M. A.; COMELLI, R. N.; SELUY, L. G. Wastewater from the soft drinks industry as a source for bioethanol production. Bioresource Technology, v. 136, p. 140–147, 2013; MAX, B. et al. Biotechnological production of citric acid. Brazilian Journal of Microbiology, v. 41, n. 4, p. 862–875, 2010)
[010] . US9677036B1 descreve uma unidade de produção de etanol que utiliza como substrato diversas matérias-primas fermentáveis à base de açúcar e amido. Dentre as diversas matérias-primas, os autores reivindicam a utilização de refrigerantes, bebidas energéticas e sucos de frutas no processo. A obtenção de etanol para utilização como combustível para veículos envolve as etapas de fermentação e destilação.[010] . US9677036B1 describes an ethanol production unit that uses as substrate various fermentable raw materials based on sugar and starch. Among the various raw materials, the authors claim the use of soft drinks, energy drinks and fruit juices in the process. Obtaining ethanol for use as fuel for vehicles involves the steps of fermentation and distillation.
[011] . A pesquisa em diversos bancos de patentes não apresentou informações referentes à utilização de descartes da indústria de bebidas açucaradas para a produção de ácido cítrico.[011] . The search in several patent banks did not provide information regarding the use of waste from the sugary beverage industry for the production of citric acid.
[012] . A presente invenção tem como objetivo a produção de ácido cítrico a partir da fermentação submersa de descartes da indústria de bebidas açucaradas por fungos filamentosos. Além de contemplar o aproveitamento de descartes da indústria de bebidas, a presente invenção não requer etapas de pré-tratamento do substrato.[012] . The present invention aims to produce citric acid from the submerged fermentation of sugary beverage industry waste by filamentous fungi. In addition to contemplating the use of waste from the beverage industry, the present invention does not require pre-treatment steps for the substrate.
[013] . Conforme evidenciado no estado da técnica, a maioria dos substratos utilizados para a produção biológica de ácido cítrico necessitam de pré-tratamento para remoção de componentes inibidores e/ou transformação dos açúcares complexos em açúcares fermentáveis. Dentre os principais componentes que, em excesso, podem prejudicar o acúmulo de ácido cítrico, destacam-se os íons metálicos ferro, manganês, cálcio, magnésio e zinco.[013] . As evidenced in the state of the art, most substrates used for the biological production of citric acid require pretreatment to remove inhibitory components and/or transform complex sugars into fermentable sugars. Among the main components that, in excess, can impair the accumulation of citric acid, the metal ions iron, manganese, calcium, magnesium and zinc stand out.
[014] . A presente invenção se diferencia de outros processos por utilizar substratos com baixas concentrações de íons metálicos e alto teor de açúcares fermentáveis, não havendo necessidade de prétratamento para uso como substrato em fermentações para produção de ácido cítrico. Além disso, a invenção oferece uma alternativa economicamente e ambientalmente interessante para valorização de descartes da indústria de bebidas açucaradas, uma vez que, além de reduzir a alta carga orgânica dos mesmos, há geração de um produto de interesse comercial.[014] . The present invention differs from other processes in that it uses substrates with low concentrations of metal ions and high levels of fermentable sugars, without the need for pre-treatment for use as a substrate in fermentations for the production of citric acid. In addition, the invention offers an economically and environmentally interesting alternative for the recovery of waste from the sugary beverage industry, since, in addition to reducing the high organic load of the same, it generates a product of commercial interest.
[015] . A presente invenção apresenta um PROCESSO DE PRODUÇÃO DE ÁCIDO CÍTRICO A PARTIR DE DESCARTES DA INDÚSTRIA DE BEBIDAS AÇUCARADAS compreendendo as seguintes etapas:
- (a)Preparo do meio de cultivo
- (b)Suplementação
- (c)Inoculação
- (d)Fermentação
- (a) Preparation of the culture medium
- (b) Supplementation
- (c) Inoculation
- (d) Fermentation
[016] . A cepa utilizada no processo é cultivada e armazenada em tubos inclinados contendo meio ágar-batata-dextrose. A incubação se dá em um período de 4 a 9 dias em estufa na faixa de temperatura de 26 a 32 °C. Após o completo desenvolvimento dos esporos, os tubos são mantidos sob refrigeração. O preparo do inóculo para utilização na fermentação se dá pelo cultivo dos microrganismos em frascos Erlenmeyer contendo meio ágar-batata-dextrose, inoculados pela técnica de profundidade e incubados a 26 – 32 ºC por um período de 4 a 9 dias. Em seguida, os esporos são extraídos com uma solução de Tween 80 com concentração entre 0,01 e 0,1% (m/v) sob agitação magnética por 5 a 15 minutos. A suspensão de esporos obtida é submetida à contagem em câmara de Neubauer, para a determinação do volume de inóculo a ser adicionado aos cultivos.[016] . The strain used in the process is cultivated and stored in inclined tubes containing agar-potato-dextrose medium. Incubation takes place over a period of 4 to 9 days in an oven at a temperature range of 26 to 32 °C. After the complete development of the spores, the tubes are kept under refrigeration. The preparation of the inoculum for use in the fermentation is carried out by the cultivation of microorganisms in Erlenmeyer flasks containing agar-potato-dextrose medium, inoculated by the depth technique and incubated at 26 - 32 ºC for a period of 4 to 9 days. Then, the spores are extracted with a Tween 80 solution with a concentration between 0.01 and 0.1% (m/v) under magnetic stirring for 5 to 15 minutes. The spore suspension obtained is subjected to counting in a Neubauer chamber to determine the volume of inoculum to be added to the cultures.
[017] . Na etapa (a) o meio de cultivo líquido é composto essencialmente por carboidratos e sais. A fonte de carboidratos é constituída basicamente por um ou mais descartes da indústria de bebidas açucaradas. O descarte da indústria de bebidas açucaradas compreende uma bebida açucarada descartada durante o processamento ou cuja validade expirou e cujo teor de açúcar seja superior a 6% (m/v).[017] . In step (a) the liquid culture medium is essentially composed of carbohydrates and salts. The carbohydrate source is basically constituted by one or more wastes from the sugary beverage industry. Disposal from the sugary beverage industry comprises a sugary beverage discarded during processing or whose shelf life has expired and whose sugar content is greater than 6% (m/v).
[018] . Na etapa (a) o descarte da indústria de bebidas açucaradas compreende uma bebida açucarada composta majoritariamente por açúcares facilmente fermentáveis, tais como glicose, frutose, sacarose e maltose, cujo teor de açúcar seja superior a 6% (m/v), com baixas concentrações de íons metálicos e pH na faixa de 2,5 a 5,0, tais como refrigerantes, sucos de frutas, néctar de frutas, energéticos e bebidas esportivas isotônicas.[018] . In step (a) the disposal of the sugary beverage industry comprises a sugary beverage composed mainly of easily fermentable sugars, such as glucose, fructose, sucrose and maltose, whose sugar content is greater than 6% (m/v), with low metal ion concentrations and pH in the range of 2.5 to 5.0, such as soft drinks, fruit juices, fruit nectar, energy drinks and isotonic sports drinks.
[019] . Na etapa (a) o meio de cultivo deve ser preparado de modo a apresentar concentração de açúcares totais na faixa de 6% a 18% (m/v), mais preferencialmente na faixa de 8% a 16% (m/v).[019] . In step (a) the culture medium must be prepared so as to present a concentration of total sugars in the range of 6% to 18% (m/v), more preferably in the range of 8% to 16% (m/v).
[020] . Na etapa (b), a cada 100 g de açúcares totais, o meio deve ser suplementado com, em porcentagem mássica, 0,04 a 0,5 de nitrogênio, 0,05 a 3 de fosfato, 0,1 a 1 de magnésio, 0,0005 a 0,001 de ferro, 0,001 a 0,005 de zinco. Etanol ou metanol podem ser adicionados como indutores numa faixa de 1 a 5% (v/v). O pH do meio deve ser ajustado, se necessário, com NaOH ou KOH para atingir o intervalo de 3 a 8.[020] . In step (b), for every 100 g of total sugars, the medium must be supplemented with, by mass, 0.04 to 0.5 nitrogen, 0.05 to 3 phosphate, 0.1 to 1 magnesium , 0.0005 to 0.001 of iron, 0.001 to 0.005 of zinc. Ethanol or methanol can be added as inducers in a range of 1 to 5% (v/v). The pH of the medium should be adjusted, if necessary, with NaOH or KOH to reach the range of 3 to 8.
[021] . Na etapa (c) o meio de cultivo deve ser inoculado em proporção variando de 4 a 20% (v/v), de modo que a concentração inicial no meio de cultivo esteja na faixa de 104 a 109 esporos/mL, preferencialmente 106 esporos/mL. O inóculo pode consistir em uma solução de esporos conforme descrito, uma solução de células recuperadas ao final da fermentação ou em uma solução líquida de células previamente cultivadas. Fungos filamentosos já relatados como produtores de ácido cítrico pela literatura podem ser utilizados no processo descrito pela presente invenção tais como Aspergillus awamori, Aspergillus luchuensi, Aspergillus oryzae, Aspergillus welwitschiae, Penicillium janthinelum e, preferencialmente, Aspergillus niger, especialmente a cepa NRRL 599 que se encontra depositada no banco de cepas do Laboratório de Engenharia de Bioprocessos e Biotecnologia da Universidade Federal do Paraná, e está catalogada na Coleção de Culturas Tropical da Fundação André Tosello e também no ARS (NRRL) Culture Collection do Departamento de Agricultura dos Estados Unidos (USDA).[021] . In step (c) the culture medium must be inoculated in a proportion ranging from 4 to 20% (v/v), so that the initial concentration in the culture medium is in the range of 104 to 109 spores/mL, preferably 106 spores /ml The inoculum may consist of a spore solution as described, a solution of cells recovered at the end of fermentation, or a liquid solution of previously cultured cells. Filamentous fungi already reported as producing citric acid in the literature can be used in the process described by the present invention such as Aspergillus awamori, Aspergillus luchuensi, Aspergillus oryzae, Aspergillus welwitschiae, Penicillium janthinelum and, preferably, Aspergillus niger, especially the NRRL 599 strain that is deposited in the strain bank of the Bioprocess and Biotechnology Engineering Laboratory of the Federal University of Paraná, and is cataloged in the Tropical Culture Collection of the André Tosello Foundation and also in the ARS (NRRL) Culture Collection of the United States Department of Agriculture (USDA). ).
[022] . Na etapa (d) a fermentação se dá na modalidade submersa, em condições aeróbicas, sob agitação, nas condições ótimas de crescimento do A. niger (temperatura entre 25 e 37 ºC, preferencialmente 30ºC, e pH entre 2,0 e 5,0, preferencialmente 3,0) e por tempo de 96 a 216 h. A fermentação submersa do meio de cultivo pode ser realizada em biorreatores, preferencialmente, do tipo tanque agitado. As condições de cultivo podem ser temperatura entre 25 e 37ºC, aeração na faixa de 0,5 - 2,5 vvm, taxa de oxigenação entre 20 e 70% e tempo de cultivo de 96 a 216 h.[022] . In step (d) the fermentation takes place in the submerged mode, under aerobic conditions, under agitation, in the optimal conditions for the growth of A. niger (temperature between 25 and 37 ºC, preferably 30ºC, and pH between 2.0 and 5.0 , preferably 3.0) and for a time of 96 to 216 h. The submerged fermentation of the culture medium can be carried out in bioreactors, preferably of the stirred tank type. Cultivation conditions can be temperature between 25 and 37ºC, aeration in the range of 0.5 - 2.5 vvm, oxygenation rate between 20 and 70% and cultivation time of 96 to 216 h.
[023] . Na etapa (d) o pH do meio pode opcionalmente ser controlado pela adição de solução de NaOH ou KOH, de modo a manter o pH na faixa de 2,0 a 5,0, preferencialmente em 3,0[023] . In step (d) the pH of the medium can optionally be controlled by the addition of NaOH or KOH solution, in order to keep the pH in the range of 2.0 to 5.0, preferably at 3.0
[024] . Os exemplos a seguir ilustram os parâmetros de processo, a composição do meio de cultivo e os resultados de acordo com a matéria divulgada na presente invenção. Estes exemplos não pretendem incluir todos os aspectos da matéria, mas ilustrar parâmetros, composições e resultados representativos. Estes exemplos não se destinam a excluir equivalentes e variações da presente invenção e, portanto, não devem ser interpretados para limitar o escopo da invenção.[024] . The following examples illustrate process parameters, culture medium composition and results in accordance with the subject matter disclosed in the present invention. These examples are not intended to include all aspects of the matter, but to illustrate representative parameters, compositions and results. These examples are not intended to exclude equivalents and variations of the present invention and therefore should not be interpreted to limit the scope of the invention.
[025] . O meio de cultivo foi preparado com a adição de 93% (v/v) de uma bebida açucarada (refrigerante, néctar de fruta, bebida esportiva isotônica, suco de fruta ou energético) e suplementado com 0,2 - 1,0 g/L de ureia, 0,5 - 3,0 g/L de KH2PO4, 0,10 - 0,40 g/L de MgSO4, 0,0001 - 0,001 g/L de FeSO4, 0,002 - 0,005 g/L de ZnSO4 e 1 - 3% (v/v) de etanol. O meio foi inoculado com 2 mL de uma suspensão de esporos de Aspergillus niger resultando numa concentração inicial de 106 esporos/mL. A fermentação foi conduzida em frascos de Erlenmeyer de 250 mL com um volume inicial de 50 mL, incubados a 30°C, com agitação de 150 rpm durante 216 horas. O mesmo procedimento foi realizado utilizando dextrose comercial (100 g/L de açúcares totais) para fins comparativos (controle). Os resultados obtidos estão apresentados na tabela a seguir. 
Claims (5)
- (a)Preparo do meio de cultivo utilizando como a fonte principal de carboidratos descartes da indústria de bebidas açucaradas que compreendem bebidas descartadas durante o processamento ou cuja validade expirou e cujo teor de açúcares totais é superior a 6% (m/v), podendo ser escolhidos entre refrigerantes, sucos de frutas, néctar de frutas, bebidas esportivas isotônicas e energéticos ou a combinação desses, de modo que a concentração inicial de açúcares seja de 6 a 18% (m/v);
- (b)Suplementação do meio de cultivo com 0,04 a 0,5% de nitrogênio, 0,05 a 3% de fosfato, 0,1 a 1% de magnésio, 0,0005 a 0,001% de ferro, 0,001 a 0,005% de zinco (porcentagens mássicas) a cada 100 g de açúcares totais, podendo também ser adicionados etanol ou metanol numa faixa de 1 a 5% (v/v);
- (c)Inoculação com Aspergillus sp;
- (d)Fermentação submersa.
- (a) Preparation of the culture medium using as the main source of carbohydrates discarded from the sugary drinks industry, which comprise drinks discarded during processing or whose validity has expired and whose total sugar content is greater than 6% (m/v), which may be chosen among soft drinks, fruit juices, fruit nectar, isotonic sports drinks and energy drinks or a combination of these, so that the initial concentration of sugars is from 6 to 18% (m/v);
- (b) Supplementation of the culture medium with 0.04 to 0.5% nitrogen, 0.05 to 3% phosphate, 0.1 to 1% magnesium, 0.0005 to 0.001% iron, 0.001 to 0.005 % of zinc (mass percentages) per 100 g of total sugars, ethanol or methanol may also be added in a range of 1 to 5% (v/v);
- (c) Inoculation with Aspergillus sp;
- (d)Submerged fermentation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| BR102020026250-5A BR102020026250B1 (en) | 2020-12-21 | 2020-12-21 | CITRIC ACID PRODUCTION PROCESS FROM WASTE FROM THE SUGAR BEVERAGE INDUSTRY |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| BR102020026250-5A BR102020026250B1 (en) | 2020-12-21 | 2020-12-21 | CITRIC ACID PRODUCTION PROCESS FROM WASTE FROM THE SUGAR BEVERAGE INDUSTRY |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| BR102020026250A2 true BR102020026250A2 (en) | 2022-06-28 |
| BR102020026250B1 BR102020026250B1 (en) | 2024-02-20 |
Family
ID=83355963
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| BR102020026250-5A BR102020026250B1 (en) | 2020-12-21 | 2020-12-21 | CITRIC ACID PRODUCTION PROCESS FROM WASTE FROM THE SUGAR BEVERAGE INDUSTRY |
Country Status (1)
| Country | Link |
|---|---|
| BR (1) | BR102020026250B1 (en) |
-
2020
- 2020-12-21 BR BR102020026250-5A patent/BR102020026250B1/en active IP Right Grant
Also Published As
| Publication number | Publication date |
|---|---|
| BR102020026250B1 (en) | 2024-02-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Sanchez-Marroquin et al. | Agave juice, fermentation and chemical composition studies of some species | |
| Takahashi et al. | Effects of acetate on the growth and fermentation performance of Escherichia coli KO11 | |
| Kawa-Rygielska et al. | Utilization of concentrate after membrane filtration of sugar beet thin juice for ethanol production | |
| CN102649926A (en) | Application of candida utilis to loquat wine making and making method for loquat wine | |
| BR112014014513B1 (en) | method for the production of lactate and lactic acid from a plant extract by means of fermentation | |
| Chaudhary et al. | Screening of yeast isolates from flowers for effective ethanol production | |
| CN102666861A (en) | Fermentation process at reduced pressure | |
| Gunasekaran et al. | Fermentation pattern of Zymomonas mobilis strains on different substrates—a comparative study | |
| Corbin et al. | Low-input fermentations of Agave tequilana leaf juice generate high returns on ethanol yields | |
| Bridgers et al. | Influence of juice preparations on the fermentability of sweet sorghum | |
| CN105722988B (en) | Process for microbial fermentation of sugar-containing substrates and use of hydrogen in atomic, ionic or gaseous state in such process | |
| Parajó et al. | Xylitol from wood: study of some operational strategies | |
| KR20000053547A (en) | Method for producing xylitol or d-xylulose | |
| BR102020026250A2 (en) | CITRIC ACID PRODUCTION PROCESS FROM WASTE PRODUCTS FROM THE SUGAR BEVERAGE INDUSTRY | |
| Zhang et al. | Physicochemical characterization of pineapple peel wine | |
| Leksawasdi et al. | Longan syrup and related products: processing technology and new product developments | |
| CN107164246A (en) | A kind of thermotolerant yeast bacterium and its application | |
| JP7077620B2 (en) | Method for producing glucose composition, microbial fermentation raw material and chemical product | |
| Thorat et al. | Standardization of Process Parameters for Production of Citric Acid from Mahua Flowers (Madhuca indica) by Surface Fermentation using Aspergillus niger NCIM-545 and NCIM-595 | |
| Widmer et al. | Simultaneous saccharification and fermentation of orange processing waste to ethanol using Kluyveromyces marxianus | |
| JPH0556958B2 (en) | ||
| BR102017002910A2 (en) | ethanol production by zymomonas mobilis in solid supported submerged fermentations from lignocellulosic acid hydrolyzate waste | |
| Singh Kocher et al. | Development of a fermentation process for utilization of Emblica Officinalis Gaertn (Amla) candy waste for production of natural vinegar | |
| Kareem et al. | Microbial production of organic acids | |
| CN101096643B (en) | Method for preparing yeast with byproduct sugar solution for production of saponin by turmeric |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| B03A | Publication of a patent application or of a certificate of addition of invention [chapter 3.1 patent gazette] | ||
| B15K | Others concerning applications: alteration of classification |
Free format text: A CLASSIFICACAO ANTERIOR ERA: C12P 7/48 Ipc: C12P 7/48 (2006.01), C12N 1/14 (2006.01), C12R 1/6 |
|
| B06A | Patent application procedure suspended [chapter 6.1 patent gazette] | ||
| B09A | Decision: intention to grant [chapter 9.1 patent gazette] | ||
| B16A | Patent or certificate of addition of invention granted [chapter 16.1 patent gazette] |
Free format text: PRAZO DE VALIDADE: 20 (VINTE) ANOS CONTADOS A PARTIR DE 21/12/2020, OBSERVADAS AS CONDICOES LEGAIS |
|
| B22L | Other matters related to patents and certificates of addition of invention: notification of licence offer (art 64 par 1 of lpi) |
Free format text: CONDICOES CONTRATUAIS: 1) ROYALTIES: 5% (CINCO POR CENTO) DO PRECO DE VENDA, LIVRE DE IMPOSTOS; 2) PRAZO: ATE O TERMINO DE VIGENCIA DA PATENTE EM 21.12.2040; 3) CONDICOES DE PAGAMENTO: ANUAL, EM FUNCAO DAS VENDAS APURADAS NO PERIODO; 4) DISPONIBILIDADE DE ?KNOW-HOW?: SIM; 5) ASSISTENCIA TECNICA: NAO. - OBS: CONSULTA A CARTA PATENTE PODERA SER FEITA ATRAVES DO ENDERECO ELETRONICO WWW.INPI.GOV.BR - NO ACESSO RAPIDO = BUSCA DE PROCESSOS = FACA LOGIN COM SENHA OU TECLE CONTINUAR = PATENTE = DIGITE O NUMERO DO PROCESSO = PESQUISAR. PARA ACESSAR, CADASTRE-SE NO PORTAL DO INPI. |