WO2003008650A1 - Procede destine a preparer du cuir au moyen de protease et procede de traitement de dechets derive du traitement du cuir au moyen dudit procede - Google Patents

Procede destine a preparer du cuir au moyen de protease et procede de traitement de dechets derive du traitement du cuir au moyen dudit procede Download PDF

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Publication number
WO2003008650A1
WO2003008650A1 PCT/KR2001/001942 KR0101942W WO03008650A1 WO 2003008650 A1 WO2003008650 A1 WO 2003008650A1 KR 0101942 W KR0101942 W KR 0101942W WO 03008650 A1 WO03008650 A1 WO 03008650A1
Authority
WO
WIPO (PCT)
Prior art keywords
protease
leather
liming
bating
deliming
Prior art date
Application number
PCT/KR2001/001942
Other languages
English (en)
Inventor
Ho-Yong Park
Kwang-Hee Son
Yong-Kook Kwon
Dong-Ha Shin
Sung-Gi Min
Original Assignee
Insect Biotech Co., Ltd.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from KR10-2001-0045205A external-priority patent/KR100441377B1/ko
Application filed by Insect Biotech Co., Ltd. filed Critical Insect Biotech Co., Ltd.
Priority to BRPI0117078-3A priority Critical patent/BR0117078B1/pt
Priority to US10/483,647 priority patent/US20040214309A1/en
Publication of WO2003008650A1 publication Critical patent/WO2003008650A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C14—SKINS; HIDES; PELTS; LEATHER
    • C14C—CHEMICAL TREATMENT OF HIDES, SKINS OR LEATHER, e.g. TANNING, IMPREGNATING, FINISHING; APPARATUS THEREFOR; COMPOSITIONS FOR TANNING
    • C14C1/00—Chemical treatment prior to tanning
    • C14C1/08—Deliming; Bating; Pickling; Degreasing
    • C—CHEMISTRY; METALLURGY
    • C14—SKINS; HIDES; PELTS; LEATHER
    • C14C—CHEMICAL TREATMENT OF HIDES, SKINS OR LEATHER, e.g. TANNING, IMPREGNATING, FINISHING; APPARATUS THEREFOR; COMPOSITIONS FOR TANNING
    • C14C1/00—Chemical treatment prior to tanning
    • C14C1/06—Facilitating unhairing, e.g. by painting, by liming
    • C14C1/065—Enzymatic unhairing

Definitions

  • the present invention relates to a method for preparing leather using protease HY-3 and a method for treating wastes derived from leather processing using the same. More particularly, the present invention is concerned with a method for preparing leather, in which protease HY-3 produced from Aranicola proteolyticus HY-3 strain is added to the steps of soaking, liming, deliming and bating; and a method for treating wastewater and solid wastes derived from soaking and liming steps, and solid wastes derived from liming, deliming, bating and finishing steps, using the protease HY-3, having advantages of preparing leather of excellent quality, and environmentally friendly treatment or recycling of wastes.
  • the leather industry as raw skins processing industry requiring expensive equipment and technique, is the chief industry determining quality of leather articles, such as shoes, bags, clothes and belts.
  • leather preparation processes using physicochemical and biological procedures are classified into water-requiring wet processes and dry processes.
  • the wet process comprises the steps of brine curing for preventing putrefaction of raw skins by bacteria or molds, soaking for removing needless components of raw skins, liming for producing limed pelt, a deliming for removing lime, bating for biologically treating unnecessary components, pickling for decreasing pH, neutralization, retanning, dyeing and fatliquoring.
  • a hair saving method which is an environmentally friendly leather preparation method, uses a proteolytic enzyme for the liming and deliming steps.
  • the liming step among leather preparation processes, largely affects COD (chemical oxygen demand) .
  • the leather industry is a representative pollution generating industry because of causing water pollution and soil pollution by large quantities of wastewater and solid wastes. Most chemicals used in each step are discharged as wastewater after their use in wet processes.
  • Fig. 1 is a schematic block diagram showing a preparation process of leather, capable of using protease HY-3 of the present invention.
  • Fig. 2 is a graph showing protein amounts eluted from leather upon use of protease HY-3 of the present invention; # : control H : supernatant A : concentrated supernatant
  • the present invention pertains to a method for preparing leather by use of protease HY-3 produced from Aranicola proteolyticus HY-3 strain.
  • the method of the present invention comprises the steps of soaking, liming, deliming and bating (see Fig. 1).
  • the protease HY-3 used for the leather preparation method of the present invention microorganisms are separated from Aranicola proteolyticus HY-3 strains- cultured medium and thus the remaining enzyme-containing supernatant is used, or a mixture of the protease HY-3 and an additive for increasing stability of the enzyme or a material introduced to each step, formulated in a preparation, is used.
  • the protease HY-3 of the present invention is preferably added at an amount of 0.1-15 wt% to the steps of soaking, liming, deliming and bating, and is the microorganism-removed enzyme-containing liquid from microorganism culture medium, or may be the preparation mixed with an additive for increasing stability of the enzyme or a material introduced to each step.
  • the present protease HY-3 has a maximum activity at 37 °C with a relative activity of 75 % or more at 20-40 °C.
  • the protease shows a maximum activity at pH 8.0 and a relative activity of 80 % or more at pH 7.0-9.5
  • protease HY-3 soaking, liming and bating steps are carried out at 25-35 °C and at pH 8-9, so that proteins in the skins can be stably decomposed using the protease HY-3. Also, when large quantities of salts are added to the steps of soaking, liming, deliming and bating, most proteolytic enzymes are decreased in their activity, whereas the present protease HY-3 maintains its activity, even at high salinity of 10 %, and thus can be applied to each step. By using the protease HY-3, conventionally used organic and inorganic chemicals for leather preparation can be drastically decreased in their amounts, and thus environmentally neutral leather-processing can be performed. Hence, such protease HY-3 can be applied to leather processing, and in particular, be useful in removal of epidermis, hairs and soluble proteins through the steps of soaking, liming, deliming and bating.
  • the soaking step when animal skins or hides subjected to salting treatment are delivered to a leather factory, water should be sufficiently added in a paddle or drum for removing various dirt and unnecessary components, after which water absorbed into skins tissues over a long period of time allows the skins tissues to be restored to the normal skins softness of a live animal. So, unnecessary soluble proteins, dirt and hairs attached to raw skins or hides can be decomposed and then removed by treatment of the protease HY-3. Through such procedure, the skin tissues become smooth and a next liming step can be easily conducted.
  • the protease HY-3 can decompose cells of stratum germinativum or base cells of hair root, using a principle of decomposing not the cortex but the medulla of hairs. Additionally, hair roots and hair follicles are removed, and thus the trichopore is certainly swelled, and then hairs are eliminated, so that lime and other chemicals can be easily penetrated and thus treatment amounts of chemicals can be reduced.
  • the protease HY-3 removes unnecessary proteins in raw skins and loosens skin tissues, whereby chemicals for leather processing to be supplied after the bating step can be readily penetrated into the tissues and a bonding strength between chemicals and tissues is increased.
  • the wastes may be formed in liquid or solid state, and also be recycled.
  • protease HY-3 applicable to the leather preparation of the present invention, use is made of a microorganism culture medium, an enzyme-containing liquid remaining after the microorganism is separated from the above medium, or a formulated preparation mixed with an additive for increasing stability of the enzyme or a material introduced to each step.
  • one selected from the group consisting of a lipase and an amylase is used, together with the protease HY-3.
  • the protease HY-3 producing microorganism medium, the enzyme-containing liquid remaining after microorganism is separated from the above medium, or the formulated preparation mixed with the additive for increasing stability of the enzyme or the material introduced to each step is added to wastewater and solid wastes generated during the soaking and liming steps, solid wastes generated during the bating step, and solid wastes after the finishing step.
  • the wastes generated from the leather-preparation amount to 40-70 wt% of the initial weights of raw skins or hides. Most discharged solid wastes are classified as industrial wastes and thus burned up or simply buried.
  • the solid wastes, treatable by the protease HY-3 are exemplified by fleshing scraps, trimming scraps and hairs from the raw skins, generated during the soaking and liming steps; pelt scraps, after the bating step; and skin scraps, generated from the finishing step following a final drying.
  • fleshing scraps and trimming scraps proteins and lipids are not completely separated and are present in a mixed state, in which lipid component amounts to 30-50 % of total components.
  • at least one enzyme selected from protease HY-3, lipase or amylase may be added.
  • the solid wastes such as pelt scraps which are produced from the bating step after the liming step, comprise about 40 % of total wastes generated from leather processing.
  • the pelt scrap comprises 4.0 % lipid, 1.5 % calcium, 5.5 % ash, 50-55 % water and 35-40 % protein.
  • the protease HY-3 of the present invention usable as a metallic protease, is increased in its activity when metal ions are present.
  • the activity of the protease HY-3 is increased about 1.5 times in the presence of 1 mM calcium ions, and 1.2-1.4 times in the presence of other metal ions.
  • Metal ions, such as calcium ions are present in large amounts in the pelt scrap, so that the present protease HY- 3 is effective for decomposition of protein in the pelt scraps.
  • the wastewater discharged from the leather processing facilities includes organic, inorganic and floatable matters, and is characterized in that BOD and COD in wastewater is very high and chrome, a heavy metal having high toxicity, is present.
  • the protease HY-3 By using the protease HY-3, the amounts of activators, limes, sulfides, salts, acids and chrome can be decreased.
  • protease HY-3 secreted from such microorganism can decompose the solid wastes derived from the leather processing.
  • protease HY-3 proteins in the wastes can be hydrolyzed to peptides to make foods, cosmetics and industrial products.
  • protease can be used for saline-containing wastewater generated during the soaking and liming steps, and thus proteins in the wastewater can be recycled by decomposition of protease HY- 3.
  • waste leather can be utilized as feed and edible gelatin by cleavage of polypeptide chains in collagen molecules.
  • protease HY-3 for treatment of solid leather wastes, environmental pollution is prevented and secondary products can be obtained.
  • protease HY-3 In order to produce protease HY-3, a standard medium for growth of microorganism was sterilized under high pressure at 121 °C for 20 minutes, and then protease HY-3 producing Aranicola proteolyticus HY-3 strain (KCTC 0268BP) was added in the amount of 0.1-5 vol% on the basis of the whole volumes of the medium and cultured at 25-30 °C for 25-30 hours. The medium was subjected to membrane filtration and thus supernatant was separated from the biomass. As necessary, the supernatant was concentrated 3- 10 times through 10 kDa membrane filtration.
  • KCTC 0268BP Aranicola proteolyticus HY-3 strain
  • protease HY-3 containing tubes have more eluted proteins, compared to the control, and also much more proteins are eluted by the concentrated supernatant than by the unconcentrated supernatant of microorganism-cultured medium.
  • the protease was formulated in a preparation and thus used at the bating step .
  • Lyophilized protease HY-3 of the present invention was added in the amount of 0.5-10 wt% to 40-50 % ammonium chloride (NHC1) , 40-50 % ammonium sulfate ((NH 4 ) 2 S0 4 ), 0.005-0.01 % calcium chloride (CaCl 3 ) and 0.025-0.1 % lactose.
  • the skins were subjected to deliming step, and then to bating step using different enzyme preparations, after which the skins were observed as to their surface state. In addition, the skins were subjected to tanning and finishing steps, and then the surface state was observed.
  • As the protease preparation used in the bating step for comparison use was made of Moron (Chungmu Fermentation, Korea) and Oropon K (TFL, Germany) . The experiment procedure is summarized in Table 1, below.
  • the present protease HY-3 and controls were added at the bating step. More specifically, at the deliming step, chemicals shown in the above table 1 were added for 1 hour and then 0.2 wt% lactic acid and 0.1- 15 wt% protease were added at the bating step. The bating step was carried out for 80 minutes, and then the pickling step for treating with saline matter-containing strong acid chemicals was carried out for 12 hours or longer and the chrome-containing tanning step was performed for about 12 hours, followed by drying the skins. Thereafter, the skins were observed.
  • the skins were subjected to deliming, bating and pickling steps while using the different enzyme preparations only in the bating step.
  • the skins after bating were observed and the skins of wet-blue state were analyzed.
  • the skins treated by the formulated protease HY-3 are clearer and smoother in grain and surface of perioplie corium, compared to controls . As for wet-blue grain, the skins treated by the protease HY-3 are softer on their surfaces and higher in whiteness.
  • a deliming agent is added to increase the effect of bating.
  • deliming agent the effect of protease HY-3 by such deliming agent was examined.
  • the surface of skins after ammonium sulfate ((NH 4 ) 2 S0 4 ) was used alone in the bating step was similar to that of skin surfaces after Amoron and Oropon K were used.
  • the skins were made to crust (that is, subjected to a tanning process to prepare skins ready for making leather goods (e.g., handbag and shoes)), which was then subjected to the method as in the following table 5.
  • the surface state of skins was observed and physical strength of skins was measured.
  • the protease HY-3 is more excellent in almost all properties than the imported Oropon K.
  • the protease HY-3 treated leather was 2.5-2.7 kg/mm 2 while the Oropon K treated leather was 1.1-2.4 kg/mm 2 .
  • the former was 5.1-7.3 kg/mm 2 while the latter was 4.1-5.8 kg/mm 2 .
  • the former was 40 kg/mm 2 or larger while the latter was 32-40 kg/mm 2 .
  • Oropon K treated leather was shown as 59-82 % and 3.5-4.2 %, whereas the protease HY-3 of the present invention produced leather of 60-77 % elongation percentage and 3.3-3.9 % softness.
  • protease HY-3 produced from Aranicola proteolyticus HY-3 strain (KCTC 0268BP) with that of other protease
  • the protease HY- 3 of the present invention and the imported Oropon K were used.
  • Oropon K as a pancreatic enzyme preparation, is widely used for increasing tensile strength and softness of grain and maintaining soft grain by decomposing collagen without damaging the grain of leather.
  • the activity of protease HY-3 was measured by Braun, V. & Schmitz, G., Arch, Microbiol. 1980, 124: 55-61.
  • azo-casein 0.24 g was dissolved in 10 ml of 50 mM phosphate buffer, pH 7.5, to prepare a substrate solution. 300 ⁇ l of substrate solution was mixed with 100 ⁇ l of culture medium and reacted at 37 °C for 30 minutes. To the reaction, 300 ⁇ l of 10 % trichloroacetate was added to further react at room temperature for 1 hour. The resulting reaction was centrifuged at 7,000 rpm and the pellet was separated from the supernatant. 300 ⁇ l of supernatant was added with 30 ⁇ l of 10 % sodium hydroxide and then absorbance was measured at 420 n . 1 unit of enzyme of the present invention was defined as the amount of enzyme releasing an amount of azo and casein, sufficient to increase absorbance by 1.0 after 1 minute of digestion of azo-casein test substrate, at 37 °C.
  • 1 unit of each of protease HY-3 and Oropon K was added to a protein substrate mixture (1 mg/ml) of casein, albumin, hemoglobin and keratin, and a protein substrate mixture (5 mg/ml) of collagen and elastin, and then reacted at 37 °C for 2 hours. Thereafter, using a Bradford method, the amount of protein in samples was measured (see, Table 7) . Then, 1 unit of enzyme was defined as the amount required to produce 1 ⁇ g protein equivalent from proteolytic digestion of the substrate at 37 °C for 1 minute.
  • the protease HY-3 of the present invention shows higher decomposition activity against most substrates, exclusive of hemoglobin, than Oropon K.
  • animal skins comprise structural proteins of collagen, elastin, keratin and so on, and non-structural proteins of albumin, globulin and the like.
  • the protease HY-3 can decompose casein and albumin, which are also present in skins, and in particular, has excellent decomposition activity against keratin, which is a main component of hairs, so that it consequently functions in the liming and deliming steps.
  • protease HY-3 was 4 times more active against collagen, compared to Oropon K.
  • the results are shown in Table 8, below.

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  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Treatment And Processing Of Natural Fur Or Leather (AREA)

Abstract

L'invention concerne un procédé destiné à préparer du cuir au moyen d'une protéase et un procédé destiné à traiter les déchets dérivés du traitement du cuir au moyen dudit procédé, qui sont avantageux en termes de préparation d'un cuir d'excellentes qualités, de production de déchets réduite grâce à la réduction de la quantité de produits chimiques, et de traitement ou de recyclage de déchets de façon écologique. La protéase HY-3 produite à partir de brins Aranicola proteolyticus HY-3 est ajoutée lors de la trempe, du chaulage, du déchaulage et du confitage, pendant le traitement du cuir, le cuir étant ainsi préparé. En outre, la protéase HY-3 est ajoutée aux eaux usées et aux déchets solides générés durant le trempage et le chaulage, et des déchets solides générés durant le chaulage, le déchaulage, le confitage et le finissage, de manière que les déchets soient traités.
PCT/KR2001/001942 2001-07-14 2001-11-14 Procede destine a preparer du cuir au moyen de protease et procede de traitement de dechets derive du traitement du cuir au moyen dudit procede WO2003008650A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
BRPI0117078-3A BR0117078B1 (pt) 2001-07-14 2001-11-14 mÉtodo para preparaÇço de couro utilizando protease e mÉtodo para tratamento de dejetos derivados do processamento do couro utilizando o mesmo.
US10/483,647 US20040214309A1 (en) 2001-07-14 2001-11-14 Method for preparing leather using protease and method for treating wastes derived from leather processing

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
KR20010042627 2001-07-14
KR2001-42627 2001-07-14
KR10-2001-0045205A KR100441377B1 (ko) 2001-07-14 2001-07-26 단백질 분해효소를 이용한 피혁의 제조방법 및 피혁제조공정 폐기물의 처리방법
KR2001-45205 2001-07-26

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2008522664A (ja) * 2004-12-07 2008-07-03 アリザント ヘルスケア インク． 変動透過性を備えた暖房装置
US8535927B1 (en) 2003-11-19 2013-09-17 Danisco Us Inc. Micrococcineae serine protease polypeptides and compositions thereof
US8865449B2 (en) 2003-11-19 2014-10-21 Danisco Us Inc. Multiple mutation variants of serine protease
CN115323081A (zh) * 2022-09-23 2022-11-11 四川大学 一种防止皮革伤面和松面的软化方法

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100811744B1 (ko) * 2006-12-28 2008-03-11 주식회사 인섹트 바이오텍 아라자임을 유효성분으로 하는 암 예방 및 치료용 약학적조성물
CN106282143A (zh) * 2015-06-12 2017-01-04 深圳市大地康恩生物科技有限公司 一种皮革软化复合酶
EP3425069B1 (fr) * 2017-07-06 2020-04-15 Stahl International B.V. Procédé de déchaulage

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH03124800A (ja) * 1989-10-06 1991-05-28 Showa Denko Kk 皮革様表面層形成剤
JPH07118700A (ja) * 1993-10-20 1995-05-09 Kooken Kagaku Kk 装飾天然皮革の製造法
WO1996011285A1 (fr) * 1994-10-07 1996-04-18 Novo Nordisk A/S Procede de tannerie par traitement des cuirs et peaux comportant un traitement enzymatique des cuirs et peaux au moyen d'une protease microbienne a action trypsine

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH03124800A (ja) * 1989-10-06 1991-05-28 Showa Denko Kk 皮革様表面層形成剤
JPH07118700A (ja) * 1993-10-20 1995-05-09 Kooken Kagaku Kk 装飾天然皮革の製造法
WO1996011285A1 (fr) * 1994-10-07 1996-04-18 Novo Nordisk A/S Procede de tannerie par traitement des cuirs et peaux comportant un traitement enzymatique des cuirs et peaux au moyen d'une protease microbienne a action trypsine

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8535927B1 (en) 2003-11-19 2013-09-17 Danisco Us Inc. Micrococcineae serine protease polypeptides and compositions thereof
US8865449B2 (en) 2003-11-19 2014-10-21 Danisco Us Inc. Multiple mutation variants of serine protease
JP2008522664A (ja) * 2004-12-07 2008-07-03 アリザント ヘルスケア インク． 変動透過性を備えた暖房装置
CN115323081A (zh) * 2022-09-23 2022-11-11 四川大学 一种防止皮革伤面和松面的软化方法
CN115323081B (zh) * 2022-09-23 2023-08-18 四川大学 一种防止皮革伤面和松面的软化方法

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BR0117078B1 (pt) 2011-06-14
CN1245523C (zh) 2006-03-15
CN1529761A (zh) 2004-09-15
BR0117078A (pt) 2004-08-03

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