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Article ; Online: Enhancement of gluconic acid production by Aspergillus niger from by‐products as glucose source using pressurized air conditions

Fernandes, Sílvia / Dias, Bruna / Belo, Isabel / Lopes, Marlene

Journal of Chemical Technology & Biotechnology. 2023 Sept., v. 98, no. 9 p.2146-2153

2023

Abstract: BACKGROUND: The profitable production of gluconic acid (GA), an organic acid extensively used in the food and beverages industries, relies on the use of cheap and available substrates with high sugar content. The high oxygen demand for the microbial ... ...

Abstract	BACKGROUND: The profitable production of gluconic acid (GA), an organic acid extensively used in the food and beverages industries, relies on the use of cheap and available substrates with high sugar content. The high oxygen demand for the microbial conversion of glucose to GA is still a bottleneck of industrial production in conventional stirred‐tank reactors. Therefore, in this study, for the first time, the effect of increased air pressure up to 4 bar on GA production and sugar consumption in batch and step‐wise fed‐batch cultures in sugarcane molasses (ScM) and grape must (GM) medium was studied. RESULTS: ScM and GM, by‐products of the agro‐food industry, proved to be adequate feedstocks to replace pure glucose in GA batch production by Aspergillus niger MUM 92.13. A threefold improvement in GA productivity was obtained in batch experiments carried out at 4 bar with ScM and GM compared to those at 1 bar. Furthermore, the addition of two pulses of ScM, under pressurized conditions, increased the GA production, reaching a final concentration of 140 g L⁻¹. CONCLUSION: This work proved that the use of total air pressure up to 4 bar is an effective approach to enhance GA productivity using ScM and GM as substrates in batch and step‐wise fed‐batch cultures. Also, this study provides a novel strategy for developing a biotechnological bioprocess for GA production by A. niger from low‐cost and sugar‐rich by‐products performed in pressurized stirred tank reactors. © 2023 Society of Chemical Industry (SCI).
Keywords	Aspergillus niger ; air ; atmospheric pressure ; bioprocessing ; biotechnology ; feedstocks ; food industry ; gluconic acid ; glucose ; grape must ; molasses ; oxygen ; sugar content ; sugarcane
Language	English
Dates of publication	2023-09
Size	p. 2146-2153.
Publishing place	John Wiley & Sons, Ltd.
Document type	Article ; Online
Note	JOURNAL ARTICLE
ZDB-ID	1479465-2
ISSN	1097-4660 ; 0268-2575
ISSN (online)	1097-4660
ISSN	0268-2575
DOI	10.1002/jctb.7428
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Exploring the use of hexadecane by Yarrowia lipolytica: Effect of dissolved oxygen and medium supplementation.

Miranda, Sílvia M / Lopes, Marlene / Belo, Isabel

Journal of biotechnology

2023 Volume 380, Page(s) 29–37

Abstract: This work aimed to evaluate the effect of medium composition and volumetric oxygen transfer coefficient ( ... ...

Abstract	This work aimed to evaluate the effect of medium composition and volumetric oxygen transfer coefficient (k
MeSH term(s)	Animals ; Fatty Acids/metabolism ; Yarrowia/metabolism ; Alkanes/metabolism ; Bioreactors
Chemical Substances	Fatty Acids ; n-hexadecane (F8Z00SHP6Q) ; Alkanes
Language	English
Publishing date	2023-12-19
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	843647-2
ISSN	1873-4863 ; 0168-1656 ; 1389-0352
ISSN (online)	1873-4863
ISSN	0168-1656 ; 1389-0352
DOI	10.1016/j.jbiotec.2023.12.006
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Article ; Online: Yarrowia lipolytica produces lipid-rich biomass in medium mimicking lignocellulosic biomass hydrolysate.

Dias, Bruna / Fernandes, Helena / Lopes, Marlene / Belo, Isabel

Applied microbiology and biotechnology

2023 Volume 107, Issue 12, Page(s) 3925–3937

Abstract: In recent years, lignocellulosic biomass has become an attractive low-cost raw material for microbial bioprocesses aiming the production of biofuels and other valuable chemicals. However, these feedstocks require preliminary pretreatments to increase ... ...

Abstract	In recent years, lignocellulosic biomass has become an attractive low-cost raw material for microbial bioprocesses aiming the production of biofuels and other valuable chemicals. However, these feedstocks require preliminary pretreatments to increase their utilization by microorganisms, which may lead to the formation of various compounds (acetic acid, formic acid, furfural, 5-hydroxymethylfurfural, p-coumaric acid, vanillin, or benzoic acid) with antimicrobial activity. Batch cultures in microplate wells demonstrated the ability of Yarrowia strains (three of Y. lipolytica and one of Y. divulgata) to grow in media containing each one of these compounds. Cellular growth of Yarrowia lipolytica W29 and NCYC 2904 (chosen strains) was proven in Erlenmeyer flasks and bioreactor experiments where an accumulation of intracellular lipids was also observed in culture medium mimicking lignocellulosic biomass hydrolysate containing glucose, xylose, acetic acid, formic acid, furfural, and 5-HMF. Lipid contents of 35% (w/w) and 42% (w/w) were obtained in bioreactor batch cultures with Y. lipolytica W29 and NCYC 2904, respectively, showing the potential of this oleaginous yeast to use lignocellulosic biomass hydrolysates as feedstock for obtaining valuable compounds, such as microbial lipids that have many industrial applications. KEY POINTS: • Yarrowia strains tolerate compounds found in lignocellulosic biomass hydrolysate • Y. lipolytica consumed compounds found in lignocellulosic biomass hydrolysate • 42% (w/w) of microbial lipids was attained in bioreactor batch cultures.
MeSH term(s)	Yarrowia ; Biomass ; Furaldehyde ; Lipids
Chemical Substances	lignocellulose (11132-73-3) ; formic acid (0YIW783RG1) ; Furaldehyde (DJ1HGI319P) ; Lipids
Language	English
Publishing date	2023-05-16
Publishing country	Germany
Document type	Journal Article
ZDB-ID	392453-1
ISSN	1432-0614 ; 0171-1741 ; 0175-7598
ISSN (online)	1432-0614
ISSN	0171-1741 ; 0175-7598
DOI	10.1007/s00253-023-12565-6
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Article: Rice Husk, Brewer's Spent Grain, and Vine Shoot Trimmings as Raw Materials for Sustainable Enzyme Production.

Guimarães, Ana / Mota, Ana C / Pereira, Ana S / Fernandes, Ana M / Lopes, Marlene / Belo, Isabel

Materials (Basel, Switzerland)

2024 Volume 17, Issue 4

Abstract: Solid by-products with lignocellulosic structures are considered appropriate substrates for solid-state fermentation (SSF) to produce enzymes with diverse industrial applications. In this work, brewer's spent grain (BSG), rice husk (RH), and vine shoot ... ...

Abstract	Solid by-products with lignocellulosic structures are considered appropriate substrates for solid-state fermentation (SSF) to produce enzymes with diverse industrial applications. In this work, brewer's spent grain (BSG), rice husk (RH), and vine shoot trimmings (VSTs) were employed as substrates in SSF with
Language	English
Publishing date	2024-02-17
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2487261-1
ISSN	1996-1944
ISSN	1996-1944
DOI	10.3390/ma17040935
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Use of Pressurized and Airlift Bioreactors for Citric Acid Production by Yarrowia lipolytica from Crude Glycerol

Ferreira, Patricia / Lopes, Marlene / Belo, Isabel

Fermentation. 2022 Dec. 02, v. 8, no. 12

2022

Abstract: Citric acid production is generally carried out in an aqueous medium in stirred tank reactors (STR), where the solubility of oxygen is low and the oxygen demand of microbial cultures is high. Thus, for this bioprocess, providing adequate oxygen mass ... ...

Abstract	Citric acid production is generally carried out in an aqueous medium in stirred tank reactors (STR), where the solubility of oxygen is low and the oxygen demand of microbial cultures is high. Thus, for this bioprocess, providing adequate oxygen mass transfer rate (OTR) from the gas phase into the aqueous culture medium is the main challenge of bioreactor selection and operation. In this study, citric acid production by Yarrowia lipolytica W29 from crude glycerol, in batch cultures, was performed in two non-conventional bioreactors normally associated with high mass transfer efficiency: a pressurized STR and an airlift bioreactor. Increased OTR was obtained by raising the total air pressure in the pressurized STR and by increasing the aeration rate in the airlift bioreactor. An improvement of 40% in maximum citric acid titer was obtained by raising the air pressure from 1 bar to 2 bar, whereas, in the airlift bioreactor, a 30% improvement was attained by increasing the aeration rate from 1 vvm to 1.5 vvm. Both bioreactor types can be successfully applied for the citric acid production process using alternative ways of improving OTR than increasing mechanical stirring power input, thus leading to important operating saving costs.
Keywords	Yarrowia lipolytica ; aeration ; atmospheric pressure ; bioprocessing ; bioreactors ; citric acid ; culture media ; fermentation ; glycerol ; mass transfer ; oxygen ; solubility
Language	English
Dates of publication	2022-1202
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article ; Online
ZDB-ID	2813985-9
ISSN	2311-5637
ISSN	2311-5637
DOI	10.3390/fermentation8120700
Database	NAL-Catalogue (AGRICOLA)

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Article: Enhancing Antioxidants Extraction from Agro-Industrial By-Products by Enzymatic Treatment.

Leite, Paulina / Belo, Isabel / Salgado, José Manuel

Foods (Basel, Switzerland)

2022 Volume 11, Issue 22

Abstract: Nowadays, agro-industrial by-products are of increasing interest as a source of antioxidant compounds. Thus, alternative green techniques to extract antioxidant compounds have been pursued. The use of enzymes to release bioactive compounds through ... ...

Abstract	Nowadays, agro-industrial by-products are of increasing interest as a source of antioxidant compounds. Thus, alternative green techniques to extract antioxidant compounds have been pursued. The use of enzymes to release bioactive compounds through antioxidant activity reduces the environmental impact caused by traditional extraction systems using organic solvents. A crude enzymatic extract containing carbohydrolases was produced by solid-state fermentation (SSF) of an olive pomace and brewery spent-grain combination. The crude extract was evaluated at different temperatures and pH values and its thermostability was studied. Results showed that β-glucosidase and cellulase were more stable than xylanase, particularly cellulase, which kept 91% of its activity for 72 h at 45 °C. The extract was also applied in enzymatic treatments (ET) to liberate antioxidant compounds from winery, olive mill and brewery by-products under optimal conditions for enzymatic activities. The highest antioxidant activity was found in extracts obtained after enzymatic treatment of exhausted olive pomace (EOP). Enzymatic crude extract produced by SSF was successfully applied in the extraction of antioxidant compounds from winery, olive mill and brewery by-products. Thus, integrating SSF and enzymatic technologies is a valuable approach to implement circular economy practices in the agro-food industry.
Language	English
Publishing date	2022-11-18
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2704223-6
ISSN	2304-8158
ISSN	2304-8158
DOI	10.3390/foods11223715
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Solid State and Semi-Solid Fermentations of Olive and Sunflower Cakes with Yarrowia lipolytica: Impact of Biological and Physical Pretreatments

Costa, Ana Rita / Fernandes, Helena / Salgado, José Manuel / Belo, Isabel

Fermentation. 2023 Aug. 06, v. 9, no. 8

2023

Abstract: Lignocellulosic biomass is a promising feedstock for added value compound production in biotechnological processes such as solid-state fermentation (SSF). Although these solid materials can be directly used as substrates in fermentations in a solid state, ...

Abstract	Lignocellulosic biomass is a promising feedstock for added value compound production in biotechnological processes such as solid-state fermentation (SSF). Although these solid materials can be directly used as substrates in fermentations in a solid state, a pretreatment is often required, especially if the microorganism selected is unable to produce lignocellulosic enzymes. In the present work, several pretreatment strategies were applied to a 50% (w/w) mixture of olive and sunflower cakes before SSF for lipase production by the oleaginous yeast Yarrowia lipolytica W29. Co-culture strategies with Y. lipolytica and Aspergillus niger did not improve lipase production by the oleaginous yeast. Biological pretreatment with a fungal enzymatic extract led to a significant increase in sugar availability in the substrate mixture after a short incubation period, improving yeast growth. Microwave and ultrasound were the physical pretreatments selected and microwave irradiation proved to be the best method, resulting in 44% and 17% increases in yeast growth and lipase production, respectively, compared to the untreated mixture. An improvement in lipase activity was also observed after ultrasonic treatment in semi-solid fermentations, leading to a 2-fold increase in this enzyme activity compared to the control. The utilization of pretreatments before SSF with Y. lipolytica can increase sugars availability and result in structural changes in the solid substrate, which can improve the bioprocesses’ productivity.
Keywords	Aspergillus niger ; Helianthus annuus ; Yarrowia lipolytica ; biomass ; carboxylic ester hydrolases ; coculture ; enzyme activity ; feedstocks ; lignocellulose ; microwave radiation ; olives ; solid state fermentation ; sugars ; ultrasonic treatment ; ultrasonics ; yeasts
Language	English
Dates of publication	2023-0806
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article ; Online
ZDB-ID	2813985-9
ISSN	2311-5637
ISSN	2311-5637
DOI	10.3390/fermentation9080734
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Bioprocessing of oilseed cakes by fungi consortia: Impact of enzymes produced on antioxidants release

Souza, Daniel / Salgado, José Manuel / Cambra-López, Maria / Dias, Alberto / Belo, Isabel

Journal of Biotechnology. 2023 Feb., v. 364 p.5-12

2023

Abstract: Oilseed cakes (OC) present high potential as feedstock for the biobased industry. Biotechnological processes allow OC valorization by the production of diverse value-added products and simultaneously altering OC structure, improving their nutritional ... ...

Abstract	Oilseed cakes (OC) present high potential as feedstock for the biobased industry. Biotechnological processes allow OC valorization by the production of diverse value-added products and simultaneously altering OC structure, improving their nutritional value, and boosting OC utilization in animal feed. This work explored the use of fungi consortium of two different species as a bioprocessing approach to improve the nutritional quality of OC, obtain enzymes and antioxidants by solid-state fermentation (SSF) of sunflower cake (SFC) and rapeseed cake (RSC). Rhyzopus oryzae and Aspergillus ibericus consortium led to the highest production of cellulase (135 U/g) and β-glucosidase (265 U/g) while maximum protease (228 U/g) was obtained with A. niger and R. oryzae consortium. Maximum xylanase production (886 U/g) was observed in SSF of RSC resulting in high hemicellulose reduction. The synergistic action of lignocellulosic enzymes resulted in extracts with increased antioxidant potential with possible application as food additives against oxidative stress.
Keywords	Aspergillus ; antioxidant activity ; bioprocessing ; biotechnology ; endo-1,4-beta-glucanase ; feeds ; feedstocks ; hemicellulose ; industry ; lignocellulose ; nutritive value ; oilseeds ; oxidative stress ; proteinases ; rapeseed cake ; solid state fermentation ; sunflower cake ; synergism ; value added ; xylanases ; Co-cultures ; Filamentous fungi ; Antioxidants ; Enzymes ; Crude protein ; Fiber
Language	English
Dates of publication	2023-02
Size	p. 5-12.
Publishing place	Elsevier B.V.
Document type	Article ; Online
ZDB-ID	843647-2
ISSN	1873-4863 ; 0168-1656 ; 1389-0352
ISSN (online)	1873-4863
ISSN	0168-1656 ; 1389-0352
DOI	10.1016/j.jbiotec.2023.01.008
Database	NAL-Catalogue (AGRICOLA)

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Article: Highly aerated cultures boost gluconic acid production by the yeast-like fungus Aureobasidium pullulans

Fernandes, Sílvia / Belo, Isabel / Lopes, Marlene

Biochemical engineering journal. 2021 Nov., v. 175

2021

Abstract: The ability of Aureobasidium pullulans NCYC 4012 to produce gluconic acid (GA) was firstly studied in flask batch cultures and it was found that GA productivity was not affected by initial glucose concentration, but highest substrate uptake rate was ... ...

Abstract	The ability of Aureobasidium pullulans NCYC 4012 to produce gluconic acid (GA) was firstly studied in flask batch cultures and it was found that GA productivity was not affected by initial glucose concentration, but highest substrate uptake rate was observed at glucose concentration of 80 g L⁻¹. The effect of oxygen mass transfer, varied by changing agitation and aeration rates or total air pressure, was evaluated in batch cultures of A. pullulans in stirred tank bioreactors. Higher productivities were attained in conditions of high oxygen mass transfer and it was proven that the use of increased total pressure of 4 bar is an effective means of oxygen supply to A. pullulans cultures. At increased pressure, cells induced the antioxidant intracellular enzyme superoxide dismutase to cope with oxidative stress due to high oxygenation conditions. On the other hand, extracellular glucose oxidase activity was reduced by highly oxygenated conditions, suggesting opposite effects of oxygen on this enzyme secretion and GA production.
Keywords	Aureobasidium pullulans ; aeration ; agitation ; antioxidants ; atmospheric pressure ; bioreactors ; fungi ; gluconic acid ; glucose ; glucose oxidase ; intracellular enzymes ; mass transfer ; oxidative stress ; oxygen ; secretion ; superoxide dismutase
Language	English
Dates of publication	2021-11
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	2012139-8
ISSN	1369-703X
ISSN	1369-703X
DOI	10.1016/j.bej.2021.108133
Database	NAL-Catalogue (AGRICOLA)

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Article: Co-management of agro-industrial wastes by solid-state fermentation for the production of bioactive compounds

Leite, Paulina / Belo, Isabel / Salgado, José Manuel

Industrial crops and products. 2021 Nov. 15, v. 172

2021

Abstract: Agro-food industries face the challenge of re-using their wastes following the circular economy concept. The co-bioprocessing of wastes produced by different agro-food industries located in the same region can be a suitable strategy for the valorization ... ...

Abstract	Agro-food industries face the challenge of re-using their wastes following the circular economy concept. The co-bioprocessing of wastes produced by different agro-food industries located in the same region can be a suitable strategy for the valorization of these wastes. This work evaluated the use of mixtures of wastes from wineries, olive mill and breweries to produce lignocellulolytic enzymes and antioxidant compounds by Aspergillus niger under solid-state fermentation (SSF). The SSF of a mixture of brewery spent grain with crude and exhausted olive pomace led to the highest values of enzymatic activity (xylanase, 837 U/g, cellulase, 87 U/g; β-glucosidase, 254 U/g), being higher than enzymes activities per mass of total substrates. The highest antioxidant activity was achieved after 1 day of fermentation, being 2.3-fold higher in comparison to the unfermented wastes. SSF allowed the co-management of agro-industrial wastes from different industries.
Keywords	Aspergillus niger ; antioxidant activity ; antioxidants ; circular economy ; collaborative management ; endo-1,4-beta-glucanase ; enzyme activity ; lignocellulases ; olive pomace ; olives ; solid state fermentation ; xylanases
Language	English
Dates of publication	2021-1115
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	1132158-1
ISSN	1872-633X ; 0926-6690
ISSN (online)	1872-633X
ISSN	0926-6690
DOI	10.1016/j.indcrop.2021.113990
Database	NAL-Catalogue (AGRICOLA)

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