Mucilage samples were taken at the end of tassel emergence (BBCH 59) from each of the three rows (n = 3) using the method of Ahmed et al. (2015) (link). Maize second and third nodal roots covered in mucilage were selected (Figure 1 ). The nodal roots were cut from the stem and placed in aluminum trays. In the laboratory, soil and plant residues on the nodal roots were removed by distilled water in a coarse sieve. Thereafter, the mucilage-covered roots were submerged in distilled water until the mucilage was fully water-saturated. After 1 day, the excess water was discarded through a sieve with a mesh size of 500 μm. The hydrated mucilage was aspirated from the nodal roots with syringes. The remaining mucilage on the root tip was removed with fine forceps. The mucilage was collected in 20 ml vials, frozen at −18°C, and subsequently freeze-dried (Beta 1-8 LSCplus, Christ, Osterode, Germany).
Beta 1 8 lscplus
Manufactured by Martin Christ
Sourced in Germany
The Beta 1-8 LSCplus is a lab equipment product manufactured by Martin Christ. It is a lyophilization system designed for the freeze-drying of samples. The core function of this product is to provide a controlled environment for the freeze-drying process, which is a widely used technique in various scientific and industrial applications.
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5 protocols using beta 1 8 lscplus
1
Maize Nodal Root Mucilage Extraction
2
Vital Gluten Extraction and Characterization
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Vital gluten was produced from wheat flour (type 550, Rosenmühle GmbH, Ergolding, Germany) using a food processor with a kneading hook (MUM4405, Robert Bosch GmbH, Stuttgart, Germany), which was equipped with a perforated bowl and a polyester cloth with 80 µm mesh size. Water-soluble components of the flour were washed out with a continuous supply of desalted water. The remaining wet gluten was freeze-dried for 72 h (Beta 1-8 LSCplus, Martin Christ Gefriertrocknungsanlagen GmbH, Osterode am Harz, Germany) and then milled in an ultra-centrifugal mill (Retsch GmbH, Haan, Germany) at a constant speed of 12,000 rpm and a sieve with a mesh size of 250 µm to obtain vital gluten powder. The residual starch content of the vital gluten samples was 3.11 ± 0.16% (4 measurements, total starch enzyme kit, Megazyme International Ireland, Ltd., Wicklow, Ireland). For further measurements, the vital gluten was rehydrated following the solvent retention capacity (SRC) method (AACC 56-11.02 [71 ]). The SRC of the gluten was 133.8 ± 3.6% (59 measurements, 14% moisture basis).
3
Extraction of Bioactive Compounds from Gymnema inodorum Leaves
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The leaves of Gymnema inodorum were collected from the Chiangda Organic Garden, Chiang Mai, Thailand. Identification and authentication of the plant was done by a taxonomist at Chiang Mai University, where a voucher specimen was coded and deposited for further studies. For extraction, fresh leaves of G. inodorum were thoroughly washed with tap water and dried in a hot air oven with a temperature of 50 °C overnight. The dried powdered leaves were then prepared using an electric blender and extracted in distilled water (DW) at a proportion of 5 g%. To facilitate the extraction process, the mixture was incubated at 60 °C with a mechanical shaker for 15 mins. After centrifugation at 2500/min for 5 min, supernatant was collected and lyophilization was subsequently performed using Beta 1-8 LSCplus (Martin Christ Gefriertrocknungsanlagen GmbH, Osterode am Harz, Germany). Aqueous crude extract of G. inodorum leaves (GIE) was stored at −20 °C for further experiments. The percentage yield after freeze-drying was 17.43% (w/w).
4
Enzymatic Hydrolysis of Gelatin
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A 5% (w/v) solution was prepared from gelatin with pure water and dissolved homogeneously. Enzymatic hydrolysis was carried out with alc and sav, and its pH was adjusted to 8.0 (60 °C) and 9.0 (50 °C), respectively, with 0.1 M NaOH/HCl solution (E/S:2.5/100 [v/p]) for 3 h. After hydrolysis, the enzymes were inactivated by heat (90 °C, 20 min), and the solutions were centrifuged at 10,000 rpm for 15 min (Andreas Hettich GmbH and Co. KG, Tuttlingen, Germany). The resulting gelatin hydrolysates were dried with a freeze dryer (Martin Christ GmbH, Beta 1–8 LSCplus, Osterode am Harz, Germany), packaged, and stored.
5
Gel Formation for SEM Imaging
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To produce gels for SEM imaging the solutions were prepared according to the procedure described in Section 2.2 . For gelation, the samples were heated in a water bath at 90 °C for 30 min. After cooling, cubes with an edge length of 1 cm were cut from the gels. To observe the optical effects of the thermally induced gel formation, photos were taken after gelation. For freezing the samples were immersed in liquid nitrogen. The sample 100 mM, 5 wt%, Lev4 did not gel during the heat treatment. Therefore, it was placed in a container made of aluminum foil and frozen therein. Immediately after freezing, the samples were dehydrated by freeze drying (Beta 1–8 LSCplus, Martin Christ Gefriertrocknungsanlagen GmbH, Osterode, Germany). Lyophilized samples were carefully broken into pieces and the breakage site was gold sputtered in a sputter coater SCD 030 (Balzers, Wiesbaden-Nordenstadt, Germany). SEM imaging was carried out at the Center for Electron Microscopy (ZELMI, Technische Universität Berlin, Berlin, Germany) with an S-2700 scanning electron microscope (Hitachi, Tokyo, Japan). Images were recorded at a magnification of 100×, 300× and 1000×. SEM was carried out at least on time for each formulation.
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