Modeling enzymatic hydrolysis of lignocellulosic substrates using confocal fluorescence microscopy I: Filter paper cellulose

2014 ◽  
Vol 112 (1) ◽  
pp. 21-31 ◽  
Author(s):  
Jeremy S. Luterbacher ◽  
Jose M. Moran-Mirabal ◽  
Eric W. Burkholder ◽  
Larry P. Walker
Keyword(s):  
Enzymatic Hydrolysis ◽  
Fluorescence Microscopy ◽  
Filter Paper ◽  
Confocal Fluorescence Microscopy ◽  
Lignocellulosic Substrates ◽  
Confocal Fluorescence ◽  
Hydrolysis Of

scholarly journals Improvement of Enzymatic Saccharification of Cellulose-Containing Raw Materials Using Aspergillus niger

Processes ◽  
2021 ◽  
Vol 9 (8) ◽  
pp. 1360
Author(s):  
Ekaterina Budenkova ◽  
Stanislav Sukhikh ◽  
Svetlana Ivanova ◽  
Olga Babich ◽  
Vyacheslav Dolganyuk ◽  
...  
Keyword(s):  
Enzymatic Hydrolysis ◽  
Aspergillus Niger ◽  
Filter Paper ◽  
Raw Materials ◽  
Enzymatic Saccharification ◽  
Wood Chip ◽  
Cellulase Activity ◽  
Wood Chips ◽  
Acid Pretreatment ◽  
Hydrolysis Of

Enzymatic hydrolysis of cellulose-containing raw materials, using Aspergillus niger, were studied. Filter paper, secondary cellulose-containing or starch-containing raw materials, miscanthus cellulose after alkaline or acid pretreatment, and wood chip cellulose, were used as substrates. The study focused on a wild A. niger strain, treated, or not (control), by ultraviolet (UV) irradiations for 45, 60, or 120 min (UV45, UV60, or UV120), or by UV irradiation for 120 min followed by a chemical treatment with NaN3 + ItBr for 30 min or 80 min (UV120 + CH30 or UV120 + CH80). A mixture of all the A. niger strains (MIX) was also tested. A citrate buffer, at 50 mM, wasthe most suitable for enzymatic hydrolysis. As the UV exposure time increased to 2 h, the cellulase activity of the surviving culturewas increased (r = 0.706; p < 0.05). The enzymatic activities of the obtained strains, towards miscanthus cellulose, wood chips, and filter paper, were inferior to those obtained with commercial enzymes (8.6 versus 9.1 IU), in some cases. Under stationary hydrolysis at 37 °C, pH = 4.7, the enzymatic activity of A. niger UV120 + CH30 was 24.9 IU. The enzymatic hydrolysis of secondary raw materials, using treated A. niger strains, was themost effective at 37 °C. Similarly, the most effective treatment of miscanthus cellulose and wood chips occurred at 50 °C. The maximum conversion of cellulose to glucose was observed using miscanthus cellulose (with alkaline pretreatment), and the minimum conversion was observed when using wood chips. The greatest value of cellulase activity was evidenced in the starch-containing raw materials, indicating that A. niger can ferment not only through cellulase activity, but also via an amylolytic one.

scholarly journals Analysis of Polyhydroxyalkanoates Granules in Haloferax mediterranei by Double-Fluorescence Staining with Nile Red and SYBR Green by Confocal Fluorescence Microscopy

Polymers ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 1582
Author(s):  
Verónica Cánovas ◽  
Salvador Garcia-Chumillas ◽  
Fuensanta Monzó ◽  
Lorena Simó-Cabrera ◽  
Carmen Fernández-Ayuso ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Culture Media ◽  
Nile Red ◽  
Sybr Green ◽  
Confocal Fluorescence Microscopy ◽  
Fluorescence Staining ◽  
Haloferax Mediterranei ◽  
Optimized Protocol ◽  
Confocal Fluorescence ◽  
Pha Production

Haloferaxmediterranei is a haloarchaeon of high interest in biotechnology because it produces and mobilizes intracellular polyhydroxyalkanoate (PHA) granules during growth under stress conditions (limitation of phosphorous in the culture media), among other interesting metabolites (enzymes, carotenoids, etc.). The capability of PHA production by microbes can be monitored with the use of staining-based methods. However, the staining of haloarchaea cells is a challenging task; firstly, due to the high ionic strength of the medium, which is inappropriate for most of dyes, and secondly, due to the low permeability of the haloarchaea S-layer to macromolecules. In this work, Haloferax mediterranei is used as a halophilic archaeon model to describe an optimized protocol for the visualization and analysis of intracellular PHA granules in living cells. The method is based on double-fluorescence staining using Nile red and SYBR Green by confocal fluorescence microscopy. Thanks to this method, the capability of PHA production by new haloarchaea isolates could be easily monitored.

Novel nontoxic mitochondrial probe for confocal fluorescence microscopy

2006 ◽  
Vol 11 (3) ◽  
pp. 034014 ◽  
Author(s):  
Silvia Versari ◽  
Anna Maria Villa ◽  
Alessandro Villa ◽  
Silvia Maria Doglia ◽  
Giorgio A. Pagani ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Confocal Fluorescence Microscopy ◽  
Confocal Fluorescence
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