An Alkalothermophilic Amylopullulanase from the Yeast <i>Clavispora lusitaniae</i> ABS7: Purification, Characterization and Potential Application in Laundry Detergent
In the present study, α-amylase and pullulanase from <i>Clavispora lusitaniae</i> ABS7 isolated from wheat seeds were studied. The gel filtration and ion-exchange chromatography revealed the presence of α-amylase and pullulanase activities in the same fraction with yields of 23.88% and 2...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-11-01
|
| Series: | Catalysts |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4344/11/12/1438 |
| _version_ | 1797506320400646144 |
|---|---|
| author | Scheherazed Dakhmouche Djekrif Leila Bennamoun Fatima Zohra Kenza Labbani Amel Ait Kaki Tahar Nouadri André Pauss Zahia Meraihi Louisa Gillmann |
| author_facet | Scheherazed Dakhmouche Djekrif Leila Bennamoun Fatima Zohra Kenza Labbani Amel Ait Kaki Tahar Nouadri André Pauss Zahia Meraihi Louisa Gillmann |
| author_sort | Scheherazed Dakhmouche Djekrif |
| collection | DOAJ |
| description | In the present study, α-amylase and pullulanase from <i>Clavispora lusitaniae</i> ABS7 isolated from wheat seeds were studied. The gel filtration and ion-exchange chromatography revealed the presence of α-amylase and pullulanase activities in the same fraction with yields of 23.88% and 21.11%, respectively. SDS-PAGE showed a single band (75 kDa), which had both α-amylase (independent of Ca<sup>2+</sup>) and pullulanase (a calcium metalloenzyme) activities. The products of the enzymatic reaction on pullulan were glucose, maltose, and maltotriose, whereas the conversion of starch produced glucose and maltose. The α-amylase and pullulanase had pH optima at 9 and temperature optima at 75 and 80 °C, respectively. After heat treatment at 100 °C for 180 min, the pullulanase retained 42% of its initial activity, while α-amylase maintained only 38.6%. The cations Zn<sup>2+</sup>, Cu<sup>2+</sup>, Na<sup>+</sup>, and Mn<sup>2+</sup> increased the α-amylase activity. Other cations Hg<sup>2+</sup>, Mg<sup>2+</sup>, and Ca<sup>2+</sup> were stimulators of pullulanase. Urea and Tween 80 inhibited both enzymes, whereas EDTA only inhibited pullulanase. In addition, the amylopullulanase retained its activity in the presence of various commercial laundry detergents. The performance of the alcalothermostable enzyme of <i>Clavispora lusitaniae</i> ABS7 qualified it for the industrial use, particularly in detergents, since it had demonstrated an excellent stability and compatibility with the commercial laundry detergents. |
| first_indexed | 2024-03-10T04:30:57Z |
| format | Article |
| id | doaj.art-8afbb9b2738f4a8282542a14b9b8856e |
| institution | Directory Open Access Journal |
| issn | 2073-4344 |
| language | English |
| last_indexed | 2024-03-10T04:30:57Z |
| publishDate | 2021-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Catalysts |
| spelling | doaj.art-8afbb9b2738f4a8282542a14b9b8856e2023-11-23T04:08:58ZengMDPI AGCatalysts2073-43442021-11-011112143810.3390/catal11121438An Alkalothermophilic Amylopullulanase from the Yeast <i>Clavispora lusitaniae</i> ABS7: Purification, Characterization and Potential Application in Laundry DetergentScheherazed Dakhmouche Djekrif0Leila Bennamoun1Fatima Zohra Kenza Labbani2Amel Ait Kaki3Tahar Nouadri4André Pauss5Zahia Meraihi6Louisa Gillmann7Departement des Sciences Naturelles, Ecole Normale Supérieure-ENS-Assia Djebar, Constantine 25000, AlgeriaLaboratoire de Génie Microbiologique et Applications, Faculté des Sciences Naturelles et de la Vie, Université des Frères Mentouri, Constantine 25000, AlgeriaDepartement des Sciences Naturelles, Ecole Normale Supérieure-ENS-Assia Djebar, Constantine 25000, AlgeriaInstitut de la Nutrition, de l’Alimentation et des Technologies Agro-Alimentaires—INAATA, Université des Frères Mentouri, Constantine 25000, AlgeriaLaboratoire de Génie Microbiologique et Applications, Faculté des Sciences Naturelles et de la Vie, Université des Frères Mentouri, Constantine 25000, AlgeriaLaboratoire de Transformations Intégrées de la Matière Renouvelable, Département Génie des Procédés, Université de Technologie de Compiègne, Alliance Sorbonne Université, CEDEX, 60205 Compiègne, FranceLaboratoire de Génie Microbiologique et Applications, Faculté des Sciences Naturelles et de la Vie, Université des Frères Mentouri, Constantine 25000, AlgeriaSONAS-IUT Laboratory, University of Angers, 49016 Angers, FranceIn the present study, α-amylase and pullulanase from <i>Clavispora lusitaniae</i> ABS7 isolated from wheat seeds were studied. The gel filtration and ion-exchange chromatography revealed the presence of α-amylase and pullulanase activities in the same fraction with yields of 23.88% and 21.11%, respectively. SDS-PAGE showed a single band (75 kDa), which had both α-amylase (independent of Ca<sup>2+</sup>) and pullulanase (a calcium metalloenzyme) activities. The products of the enzymatic reaction on pullulan were glucose, maltose, and maltotriose, whereas the conversion of starch produced glucose and maltose. The α-amylase and pullulanase had pH optima at 9 and temperature optima at 75 and 80 °C, respectively. After heat treatment at 100 °C for 180 min, the pullulanase retained 42% of its initial activity, while α-amylase maintained only 38.6%. The cations Zn<sup>2+</sup>, Cu<sup>2+</sup>, Na<sup>+</sup>, and Mn<sup>2+</sup> increased the α-amylase activity. Other cations Hg<sup>2+</sup>, Mg<sup>2+</sup>, and Ca<sup>2+</sup> were stimulators of pullulanase. Urea and Tween 80 inhibited both enzymes, whereas EDTA only inhibited pullulanase. In addition, the amylopullulanase retained its activity in the presence of various commercial laundry detergents. The performance of the alcalothermostable enzyme of <i>Clavispora lusitaniae</i> ABS7 qualified it for the industrial use, particularly in detergents, since it had demonstrated an excellent stability and compatibility with the commercial laundry detergents.https://www.mdpi.com/2073-4344/11/12/1438α-amylasepullulanase<i>Clavispora lusitaniae</i>purificationenzyme characterization |
| spellingShingle | Scheherazed Dakhmouche Djekrif Leila Bennamoun Fatima Zohra Kenza Labbani Amel Ait Kaki Tahar Nouadri André Pauss Zahia Meraihi Louisa Gillmann An Alkalothermophilic Amylopullulanase from the Yeast <i>Clavispora lusitaniae</i> ABS7: Purification, Characterization and Potential Application in Laundry Detergent Catalysts α-amylase pullulanase <i>Clavispora lusitaniae</i> purification enzyme characterization |
| title | An Alkalothermophilic Amylopullulanase from the Yeast <i>Clavispora lusitaniae</i> ABS7: Purification, Characterization and Potential Application in Laundry Detergent |
| title_full | An Alkalothermophilic Amylopullulanase from the Yeast <i>Clavispora lusitaniae</i> ABS7: Purification, Characterization and Potential Application in Laundry Detergent |
| title_fullStr | An Alkalothermophilic Amylopullulanase from the Yeast <i>Clavispora lusitaniae</i> ABS7: Purification, Characterization and Potential Application in Laundry Detergent |
| title_full_unstemmed | An Alkalothermophilic Amylopullulanase from the Yeast <i>Clavispora lusitaniae</i> ABS7: Purification, Characterization and Potential Application in Laundry Detergent |
| title_short | An Alkalothermophilic Amylopullulanase from the Yeast <i>Clavispora lusitaniae</i> ABS7: Purification, Characterization and Potential Application in Laundry Detergent |
| title_sort | alkalothermophilic amylopullulanase from the yeast i clavispora lusitaniae i abs7 purification characterization and potential application in laundry detergent |
| topic | α-amylase pullulanase <i>Clavispora lusitaniae</i> purification enzyme characterization |
| url | https://www.mdpi.com/2073-4344/11/12/1438 |
| work_keys_str_mv | AT scheherazeddakhmouchedjekrif analkalothermophilicamylopullulanasefromtheyeasticlavisporalusitaniaeiabs7purificationcharacterizationandpotentialapplicationinlaundrydetergent AT leilabennamoun analkalothermophilicamylopullulanasefromtheyeasticlavisporalusitaniaeiabs7purificationcharacterizationandpotentialapplicationinlaundrydetergent AT fatimazohrakenzalabbani analkalothermophilicamylopullulanasefromtheyeasticlavisporalusitaniaeiabs7purificationcharacterizationandpotentialapplicationinlaundrydetergent AT amelaitkaki analkalothermophilicamylopullulanasefromtheyeasticlavisporalusitaniaeiabs7purificationcharacterizationandpotentialapplicationinlaundrydetergent AT taharnouadri analkalothermophilicamylopullulanasefromtheyeasticlavisporalusitaniaeiabs7purificationcharacterizationandpotentialapplicationinlaundrydetergent AT andrepauss analkalothermophilicamylopullulanasefromtheyeasticlavisporalusitaniaeiabs7purificationcharacterizationandpotentialapplicationinlaundrydetergent AT zahiameraihi analkalothermophilicamylopullulanasefromtheyeasticlavisporalusitaniaeiabs7purificationcharacterizationandpotentialapplicationinlaundrydetergent AT louisagillmann analkalothermophilicamylopullulanasefromtheyeasticlavisporalusitaniaeiabs7purificationcharacterizationandpotentialapplicationinlaundrydetergent AT scheherazeddakhmouchedjekrif alkalothermophilicamylopullulanasefromtheyeasticlavisporalusitaniaeiabs7purificationcharacterizationandpotentialapplicationinlaundrydetergent AT leilabennamoun alkalothermophilicamylopullulanasefromtheyeasticlavisporalusitaniaeiabs7purificationcharacterizationandpotentialapplicationinlaundrydetergent AT fatimazohrakenzalabbani alkalothermophilicamylopullulanasefromtheyeasticlavisporalusitaniaeiabs7purificationcharacterizationandpotentialapplicationinlaundrydetergent AT amelaitkaki alkalothermophilicamylopullulanasefromtheyeasticlavisporalusitaniaeiabs7purificationcharacterizationandpotentialapplicationinlaundrydetergent AT taharnouadri alkalothermophilicamylopullulanasefromtheyeasticlavisporalusitaniaeiabs7purificationcharacterizationandpotentialapplicationinlaundrydetergent AT andrepauss alkalothermophilicamylopullulanasefromtheyeasticlavisporalusitaniaeiabs7purificationcharacterizationandpotentialapplicationinlaundrydetergent AT zahiameraihi alkalothermophilicamylopullulanasefromtheyeasticlavisporalusitaniaeiabs7purificationcharacterizationandpotentialapplicationinlaundrydetergent AT louisagillmann alkalothermophilicamylopullulanasefromtheyeasticlavisporalusitaniaeiabs7purificationcharacterizationandpotentialapplicationinlaundrydetergent |