Patent Application
20250204522
Applicant hereby makes priority claim to a Taiwan application, number 112150386, having the Taiwan filing date of Dec. 22, 2023.
Table I (in Sequence Listing XML format) shows the at least 253 peptides and its annotated sequences for the solution generated in accordance with the disclosure of this application. The Sequence Listing XML file complies with the WIPO ST.26 requirements. Said XML copy, created on Mar. 17, 2024, is named Table-I-253_sequence and is 216 bytes in size.
Applicant hereby incorporates by reference said Sequence Listing XML file in its entirety as part of the disclosure and specification of the present application
Present invention disclosed and claimed the method and application of a keratin hydrolysis peptide (“KHP”) solution to mitigate cotton damages due to herbicides. Such mitigation is especially effective when the KHP solution is applied, either by soil infusion or by foliar spray, to the cotton plants, resulting in the reduction of damages to above-ground biomass and under-ground roots under the stress of herbicides.
Weeds are ever-present in farm lands, competing against the intended crops for light, water, nutrients and growth space. Weeds also serve as the breeding grounds of germs, vermin and rodents that feed on crops, causing economic damages greatly. During the cotton's growth cycles, weeds traditionally posed a major problem to the effective harvesting of cotton.
In the cotton growing industries, people have been using herbicides to control the growth and spreading of weeds for decades. The amount of herbicides usage needs to be accurate: excessive usage would lead to injuries to the cotton crops whereas low dosage would not provide any hindrance to the weeds' growth and development.
Also, the timing of herbicides application is critical. If the herbicide application timing is improper, there will be direct injuries to the cotton plants' development, reducing photosynthesis and other metabolisms while the weeds' growth is not affected. In other words, the herbicide's application timing should be set at a stage where the cotton plant is most resistant to the herbicides and the weeds are most vulnerable.
The inventors of present application found out that, at an early growth stage, the KHP solution made as taught herein can be applied to cotton plants, to effectively reduce and mitigate the damages caused by the herbicide and in fact contributed to better development of cotton's biomass, both underground and above-ground portions.
Hydrolyzed keratin has long been used to strengthen hairs, reduce hair splitting and breakage. Other beneficial uses include skin moisturization and wound healing. Keratin hydrolysate has also been known to function as a biofertilizer, boosting plants' growth by enhancing the plants' ability to receive and utilize nutrients, including commonly applied fertilizers.
The KHP solution is made by a hydrolysis process using feathers and water, via a high-temperature and high-pressure process, resulting in a solution that has many beneficial applications in the fields of horticulture, agriculture and potentially other farming businesses.
As disclosed herein, the application of the KHP solutions can be done by infusing to the soil of young cotton plants, or by spraying to the leaf surface of cotton plants.
The keratin solution is primarily based upon feather, which contains 85-91% keratin, 13-15% organic nitrogen, 1.6-2% organic sulfur, as well as other materials. The high keratin content has drawn many prior researches that work to break down, by enzyme, chemical agents, or fermentation process, into peptides, amino acids and other smaller molecules that can be used for animal feeds, plant fertilizers, and cultivation bases.
Around 2019, Nurdiawati, et al, came up with a hydrolysis process, by the mixture of α-amylase and protease to hydrolyze feather waste, resulting in a mixture of amino acids, fatty acids, and sugars. Nurdiawati experimented and adopted certain specific high-temperature and high-pressure setting in the hydrolysis process and discovered that the resulting solution, when mixed with some potassium and other minerals, can boost the growth of Pogostemon cablin and Vigna radiata, as reported in International Journal of Recycling or Organic Waste in Agriculture (8:221-232, 2019).
The inventors of present application, under the aegis of CH Biotech, developed and selected different feather and water compositions to perform the hydrolysis at higher temperature and higher pressure setting, resulting with different keratin hydrolysis peptide (“KHP”) solution that can be used on different crops/plants.
The selected embodiment of present invention uses a mixture of water and feathers, and subject the mixture to a thermal hydrolysis process to create KHP solutions based upon temperature/pressure parameters as noted below.
The inventors used Dionex UltiMate 3000 UPLC to separate the peptides; an analysis is done via Thermo Orbitrap Fusion Lumos Tribrid Orbitrap mass spectrometry to identify the peptides, which are then subsequently confirmed by looking up the BIOPEP-UWM database.
The application of the KHP solution can be diluted. The range of dilution ratio between 50 times and—800 times (noted as 50×-800×) of water by volume can be done, with a preferred narrower range of 50×-500×. In present application, the inventors tested, and confirmed, the specific dilution ratio of 100× and 500×.
The accompanying drawings, figures and tables, which are incorporated in and constitute a part of this specification, illustrate and exemplify the preferred embodiments of the invention. Together with the description, serve to explain the principles of the invention.
The keratin hydrolysis peptide (“KHP”) solution of present invention is made by a high-temperature and high-pressure process to treat a mixture of water and feathers as shown in the parameters herein.
The mixture ratio, temperature, pressure and duration parameters can have the three sets below:
The KHP solution in present application embodied the second set of parameters shown here and takes the steps of:
The keratin hydrolysis peptide (KHP) solution of the second embodiment is further filtered and concentrated to 301,500 ppm concentration.
The confirmation of some of the 253 peptides is further done by referencing the BIOPEP-UWM database.
The KHP solution in the selected embodiment can be diluted with water at the ratio of between 50 times to 800 times, denoted as 50×-800×, before application to the cotton plants. A preferred and narrower range is 50×-500×.
The inventors tested the KHP solutions by two methods of application to combat the adverse impact of herbicides: soil infusion and leaf spray.
For the soil infusion method of testing the mitigation effect, the KHP solution can be applied to the soil when young cotton seedlings are grown; immediately thereafter, herbicides are sprayed onto the leaves of the cotton seedlings.
For the leaf spray method of testing the mitigation effect, the KHP solution can be mixed with the herbicide in a liquid form, then sprayed to the surface of the cotton leaves.
The inventors selected the cotton species of Deltapine 1646 B2XF, to test the mitigation effective of KHP solution when the herbicide of Roundup Power Max is used.
The inventors prepared 5-inch starter pots to seed the cottons, giving them the growth environment of 16 hours day time, at 28° C., and 8 hours of night time, at of 25° C.
When the first true leaf appears, which is roughly the 22nd day after seeding, the cotton seedlings are transplanted into regular pots for experiments, where 0.13 g of HauBao No. 2 fertilizer is given to each regular pot.
The inventors divide the pots into four (4) groups:
With the four groups as defined above, the KHP solution is applied to cotton seedlings in the conditions set forth below.
For leaf spray, the Roundup Power Max, diluted 120 times with water, is thoroughly mixed with KHP_500× solution. For each cotton seedling, 2 ml of the mixed solution is sprayed on it by using a spray gun.
For soil infusion, each regular pot is infused with 100 ml of KHP_100× solution. Immediately after the infusion, 2 ml of the Roundup Power Max, diluted 120 times with water, is sprayed onto the cotton seedling.
The table below summarized the conditions of the KHP solution application:
On the 21st day after the treatment, measurements are taken from all the group to confirm the effectiveness of the application of KHP solution.
The digital scale AP224X (Shimadazu) is used to measure both the above-ground and underground biomass. The main stems and leaves are photographed and observed, using WinFOLIA Pro 2014a (Regent Instruments, Inc.) to compute and analyze leaf surface areas. The results are then tabulated into the figures as presented herein.
The stem widths from the groups are shown in
On the 21st day after the treatment, the inventors computed the surface areas from the 3rd to the 10th leaves of the main stem and tabulated the analysis results.
As shown in
The measurements of underground biomass weights are shown in
Based upon the tests and experiments done by the inventors, it is confirmed that application of the KHP solutions, at the dilution ratios and the growth stage as disclosed herein, effectively remedied the injuries to cotton plants while the goal of controlling weeds is achieved.
While the disclosure herein gave limited teachings and embodiment examples, it should be noted that the description and disclosure made herein illustrated the preferred embodiments of the invention and are not meant to limit the scope of the applicant's rights. Variations and alterations may be employed for yet additional embodiments without departing from the scope of the invention herein.
| Number | Date | Country | Kind |
|---|---|---|---|
| 112150386 | Dec 2023 | TW | national |
